Name: Human Exhausted T Cell Antibody Sampler Kit
Brand: Cell Signaling Technology

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Human Exhausted T Cell Antibody Sampler Kit: Image 1

Flow cytometric analysis of human peripheral blood mononuclear cells gated on lymphocytes co-stained with CD3 (UCHT1) Mouse mAb (APC Conjugate) #19881 using Tox/Tox2 (E6G5O) Rabbit mAb (right) compared to concentration matched

Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated SU-DHL-4 cells and either Tox/Tox2 (E6G5O) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP^® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human HERC2 intron 1 primers, human WAPAL exon 1 primers, and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Human Exhausted T Cell Antibody Sampler Kit: Image 3

Western blot analysis of extracts from various human cell lines using Tox/Tox2 (E6G5O) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from mouse thymocytes and either TCF1/TCF7 (C63D9) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse LEF1 Upstream Primers #80993 and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Human Exhausted T Cell Antibody Sampler Kit: Image 5

Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and either EOMES (D8D1R) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human PRF1 Promoter Primers #9014, human CCL4 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Human Exhausted T Cell Antibody Sampler Kit: Image 6

Flow cytometric analysis of fixed and permeabilized human peripheral blood mononuclear cells, untreated (left column) or treated with anti-CD3 (10ug/ml, 72hr) and anti-CD28 (5ug/ml, 72 hr; right column), using PD-1 (D4W2J) XP^® Rabbit mAb #86163 (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (bottom row), and co-stained with CD3 (UCHT1) Mouse mAb (APC Conjugate) #19881. Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 7

Immunohistochemical analysis of paraffin-embedded human infiltrating papillary carcinoma of the breast using PD-1 (D4W2J) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Human Exhausted T Cell Antibody Sampler Kit: Image 8

Immunoprecipitation of CTLA-4 protein from CD4⁺ T cell extracts. CD4⁺ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900, and lane 3 is CTLA-4 (E1V6T) Rabbit mAb. Western blot analysis was performed using CTLA-4 (E1V6T) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 9

Confocal immunofluorescent analysis of human CD8⁺ T cells using TIGIT (E5Y1W) XP^® Rabbit mAb (green) and CD8α (RPA-T8) Mouse mAb (FITC Conjugate) #55397 (red pseudocolor). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue). CD8⁺ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (20 ng/mL).

Human Exhausted T Cell Antibody Sampler Kit: Image 10

Immunohistochemical analysis of paraffin-embedded human breast ductal carcinoma using LAG3 (D2G4O™) XP® Rabbit mAb performed on the Leica® Bond™ Rx.

Human Exhausted T Cell Antibody Sampler Kit: Image 11

Flow cytometric analysis of Jurkat cells (blue) and primary CD4+ T cells (green) using TIM-3 (D5D5R™) XP^® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody. CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Human Exhausted T Cell Antibody Sampler Kit: Image 12

Western blot analysis of extracts from primary human CD4+ T cells and various cell lines using TIM-3 (D5D5R™) XP^® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Human Exhausted T Cell Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded renal clear cell carcinoma using TIM-3 (D5D5R™) XP^® Rabbit mAb performed on the Leica® BOND™ Rx.

Human Exhausted T Cell Antibody Sampler Kit: Image 14

Immunoprecipitation of 2B4/SLAMF4/CD244 protein from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900, and lane 3 is 2B4/SLAMF4/CD244 (D5J9D) Rabbit mAb. Western blot analysis was performed using 2B4/SLAMF4/CD244 (D5J9D) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 15

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO^® is added and emits light during enzyme catalyzed decomposition.

Human Exhausted T Cell Antibody Sampler Kit: Image 16

Flow cytometric analysis of RPMI 8226 cells (blue, negative) and SU-DHL-4 cells (green, positive) using Tox/Tox2 (E6G5O) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^®488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 17

Western blot analysis of extracts from human, mouse, and rat tissues and mouse cell lines using Tox/Tox2 (E6G5O) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 18

Flow cytometric analysis of A20 cells (blue) and EL-4 cells (green) using TCF1/TCF7 (C63D9) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 19

Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and either EOMES (D8D1R) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Sonication Chromatin IP Kit #56383. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human PRF1 Promoter Primers #9014, human CCL4 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Human Exhausted T Cell Antibody Sampler Kit: Image 20

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PD-1 (D4W2J) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 21

Western blot analysis of extracts from various human cells using CTLA-4 (E1V6T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4⁺ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Human Exhausted T Cell Antibody Sampler Kit: Image 22

Confocal immunofluorescent analysis of MJ [G11] cells (left, positive) and IGROV-1 cells (right, negative) using TIGIT (E5Y1W) XP^® Rabbit mAb (green). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Human Exhausted T Cell Antibody Sampler Kit: Image 23

Immunohistochemical analysis of paraffin-embedded HDLM-2 (left) and PC-3 (right) cell pellets on SignalSlide^® PD-L1 IHC Controls #13747 using LAG3 (D2G4O™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 24

Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing full-length human Myc/DDK-tagged TIM-3 (hTIM-3-Myc/DDK; +), using TIM-3 (D5D5R™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human tonsil using TIM-3 (D5D5R™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 26

Western blot analysis of extracts from various cell lines using 2B4/SLAMF4/CD244 (D5J9D) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 27

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human Tox protein (hTox-Myc/DDK), Myc/DDK-tagged full-length human Tox2 protein (hTox2-Myc/DDK), Myc/DDK-tagged full-length human Tox3 protein (hTox3-Myc/DDK), or Myc/DDK-tagged full-length human Tox4 protein (hTox4-Myc/DDK), using Tox/Tox2 (E6G5O) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 28

Confocal immunofluorescent analysis of DLD-1 cells using TCF1/TCF7 (C63D9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor^® 555 phalloidin (red).

Human Exhausted T Cell Antibody Sampler Kit: Image 29

Flow cytometric analysis of human peripheral blood mononuclear cells, co-stained with a CD14 antibody (left) or a CD3 antibody (right), using EOMES (D8D1R) Rabbit mAb (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype control #3900 (bottom row). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using PD-1 (D4W2J) XP^®Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 31

Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left-top) or TIGIT-transfected (right-top), MJ [G11] cell pellet (left-bottom, positive), and purified CD8⁺human peripheral blood mononuclear cell pellet (right-bottom, positive), using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 32

Immunohistochemical analysis of paraffin-embedded human colitis using LAG3 (D2G4O™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 33

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TIM-3 (D5D5R™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 34

Confocal immunofluorescent analysis of mouse thymus (left) and pancreas (right) using Tox/Tox2 (E6G5O) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Human Exhausted T Cell Antibody Sampler Kit: Image 35

Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma using TCF1/TCF7 (C63D9) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 36

Flow cytometric analysis of RPMI 8226 cells (blue) and NK-92 cells (green), using EOMES (D8D1R) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 37

Immunohistochemical analysis of paraffin-embedded 293 cell pellets, control (left) or PD-1 transfected (right), using PD-1 (D4W2J) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 38

Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 39

Immunohistochemical analysis of paraffin-embedded human tonsil using LAG3 (D2G4O™) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 40

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TIM-3 (D5D5R™) XP^® Rabbit mAb. Note staining of alveolar macrophages.

Human Exhausted T Cell Antibody Sampler Kit: Image 41

Confocal immunofluorescent analysis of rat thymus (left) and pancreas (right) using Tox/Tox2 (E6G5O) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Human Exhausted T Cell Antibody Sampler Kit: Image 42

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TCF1/TCF7 (C63D9) Rabbit mAb in the presence of control peptide (left) or TCF1/TCF7 blocking peptide #1007 (right).

Human Exhausted T Cell Antibody Sampler Kit: Image 43

Confocal immunofluorescent analysis of NK-92 cells (left, positive) and HeLa cells (right, negative) using EOMES (D8D1R) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Human Exhausted T Cell Antibody Sampler Kit: Image 44

Multiplex immunohistochemical analysis of paraffin-embedded human breast carcinoma usng PD-1 (D4W2J) XP® rabbit mAb (green), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow), CD8α (C8/144B) mouse mAb #70306 (orange), and Pan-keratin (C11) mouse mAb #4545 (cyan).

Human Exhausted T Cell Antibody Sampler Kit: Image 45

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 46

Immunohistochemical analysis of paraffin-embedded cell pellets, primary CD4+ T cells (left) and HT-29 cells (right), using TIM-3 (D5D5R™) XP^® Rabbit mAb. CD4+ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Human Exhausted T Cell Antibody Sampler Kit: Image 47

Confocal immunofluorescent analysis of mouse colon using Tox/Tox2 (E6G5O) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Human Exhausted T Cell Antibody Sampler Kit: Image 48

Immunohistochemical analysis of paraffin-embedded human tonsil using TCF1/TCF7 (C63D9) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 49

Western blot analysis of extracts from various cell lines using EOMES (D8D1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 50

Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using PD-1 (D4W2J) XP® rabbit mAb (green), B7-H3 (D9M2L) XP® rabbit mAb #14058 (red), B7-H4 (D1M8I) XP® rabbit mAb (cyan), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow), and VISTA (D1L2G™) XP® rabbit mAb #64953 (orange).

Human Exhausted T Cell Antibody Sampler Kit: Image 51

Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 52

Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using LAG3 (D2G4O™) XP® rabbit mAb (orange), PD-1 (EH33) mouse mAb #43248 (green), CD8α (C8/144B) mouse mAb #70306 (magenta), CD68 (D4B9C) XP® rabbit mAb #76437 (red), Pan-keratin (C11) mouse mAb #4545 (cyan), and TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow).

Human Exhausted T Cell Antibody Sampler Kit: Image 53

Confocal immunofluorescent analysis of SU-DHL-4 cells (left, positive) or A549 cells (right, negative) using Tox/Tox2 (E6G5O) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Human Exhausted T Cell Antibody Sampler Kit: Image 54

Western blot analysis of total cell lysates from HT29, Colo201, Jurkat and mouse thymocytes using TCF1/TCF7 (C63D9) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 55

Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the tonsil using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 56

Immunoprecipitation of Tox protein from SU-DHL-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900, and lane 3 is Tox/Tox2 (E6G5O) Rabbit mAb. Western blot analysis was performed using Tox/Tox2 (E6G5O) Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 57

Immunoprecipitation of PD-1 protein from Molt-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900, and lane 3 is PD-1 (D4W2J) XP^® Rabbit mAb. Western blot analysis was performed using PD-1

(D4W2J) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 58

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 59

Western blot analysis of extracts from HDLM-2, HuT 78, and Jurkat cells using LAG3 (D2G4O™) XP^® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Human Exhausted T Cell Antibody Sampler Kit: Image 60

Western blot analysis of extracts from human CD4⁺ T cells, MOLT-4, and Jurkat cells using PD-1 (D4W2J) XP^® Rabbit mAb (upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4⁺ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of human interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Human Exhausted T Cell Antibody Sampler Kit: Image 61

Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using TIGIT (E5Y1W) XP^® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (right).

Human Exhausted T Cell Antibody Sampler Kit: Image 62

Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 63

Immunohistochemical analysis of paraffin-embedded human tonsil using TIGIT (E5Y1W) XP^® Rabbit mAb.

Human Exhausted T Cell Antibody Sampler Kit: Image 64

Immunoprecipitation of TIGIT protein from MJ [G11] cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^®Isotype Control #3900, and lane 3 is TIGIT (E5Y1W) XP^® Rabbit mAb. Western blot analysis was performed using TIGIT (E5Y1W) XP^® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.

Human Exhausted T Cell Antibody Sampler Kit: Image 65

Western blot analysis of extracts from human CD8⁺ T cells and various human cell lines using TIGIT (E5Y1W) XP^® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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To Purchase # 81490T

製品番号	サイズ	価格	在庫
81490T	1 Kit (9 x 20 µl)

Product Includes
Related Products

Product Includes	Quantity	Applications	Reactivity	MW(kDa)	Isotype
Tox/Tox2 (E6G5O) Rabbit mAb 36778	20 µl	WB IP IF F ChIP	H M R	60-80	Rabbit IgG
TCF1/TCF7 (C63D9) Rabbit mAb 2203	20 µl	WB IP IHC IF F ChIP	H M	48, 50	Rabbit IgG
EOMES (D8D1R) Rabbit mAb 81493	20 µl	WB IF F ChIP	H	75, 85	Rabbit IgG
PD-1 (D4W2J) XP^® Rabbit mAb 86163	20 µl	WB IP IHC F	H	52-65	Rabbit IgG
CTLA-4 (E1V6T) Rabbit mAb 96399	20 µl	WB IP	H	25-30	Rabbit IgG
TIGIT (E5Y1W) XP^® Rabbit mAb 99567	20 µl	WB IP IHC IF	H	18, 30-40	Rabbit IgG
LAG3 (D2G4O™) XP^® Rabbit mAb 15372	20 µl	WB IHC	H	60-80	Rabbit IgG
TIM-3 (D5D5R™) XP^® Rabbit mAb 45208	20 µl	WB IP IHC F	H	45-70	Rabbit IgG
2B4/SLAMF4/CD244 (D5J9D) Rabbit mAb 54560	20 µl	WB IP	H M	70-120	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074	100 µl	WB			Goat

Kit Usage Information

Protocols

2203: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow, ChIP Magnetic
7074: Western Blotting
15372: Western Blotting, Immunohistochemistry (Leica® Bond™), Immunohistochemistry (Paraffin)
36778: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Immunofluorescence, Flow, ChIP Magnetic
45208: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Leica® Bond™), Immunohistochemistry (Paraffin), Flow Cytometry Live Cell Unconjugated Rabbit
54560: Western Blotting, Immunoprecipitation (Agarose)
81493: Western Blotting, Immunofluorescence, Flow, Sonication Chromatin Immunoprecipitation Protocol
86163: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Leica® Bond™), Immunohistochemistry (Paraffin), Flow
96399: Western Blotting, Immunoprecipitation (Magnetic)
99567: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), Immunofluorescence

Product Description

The Human Exhausted T Cell Antibody Sampler Kit provides an economical means of characterizing the extent of exhaustion in T cells. This kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Human Exhausted T Cell Antibody Sampler Kit detects endogenous levels of its target human protein. Tox/Tox2 (E6G5O) Rabbit mAb does not cross-react with Tox3 or Tox4 proteins, but does cross-react with an unidentified protein of 50 kDa in some murine cell extracts. TCF1/TCF7 (C63D9) Rabbit mAb does not recognize the dominant negative isoforms of TCF1/TCF7 lacking the amino-terminal β-catenin binding domain and does not cross-react with LEF1. TIGIT (E5Y1W) XP^® Rabbit mAb cross-reacts with an unidentified protein of 42 kDa in some cell extracts. 2B4/SLAMF4/CD244 (D5J9D) Rabbit mAb cross-reacts with an unidentified protein of 18 kDa in some cell extracts.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln435 of human Tox protein, Pro96 of human TCF1/TCF7 protein, Pro180 of human EOMES protein, Ala274 of human PD-1 protein, Leu95 of human CTLA-4 protein, and Asp367 of human 2B4/SLAMF4/CD244 protein. Monoclonal antibodies are produced by immunizing animals with a recombinant protein specific to the carboxy terminus of human TIGIT protein, the extracellular domain of human TIM-3 protein, and the amino terminus of human LAG3 protein.

Background

Tox, TCF1/TCF7, and EOMES play key roles in T cell development. Tox is also induced by high antigen stimulation during chronic viral infection or cancer, regulating T cell persistence and exhaustion. TCF1/TCF7 preserves the effector function of exhausted T cells during viral infection or cancer. EOMES is a key transcription factor for memory T cells and for full effector differentiation of CD8+ T cells. Expression of EOMES is induced in CD8+ T cells following viral infection and bacterial infection, and high levels of EOMES promotes T cell exhaustion. The dynamic expression of these transcription factors help characterize the extent to which a T cell is exhausted and will respond to antigen stimulation (1-5)

PD-1 (PDCD1, CD279), CTLA-4 (CD152), TIGIT (VSIG9, VSTM3), TIM-3 (HAVCR2), LAG3 (CD223), and 2B4 (SLAMF4, CD244) are immune cell co-inhibitory receptors (also known as immune checkpoints) that negatively regulate T cell function and dampen the immune response to pathogens and cancer. In addition to activated T cells, PD-1 is expressed by activated B cells and monocytes. Following interaction with its ligands, PD-L1 and PD-L2, PD-1 is phosphorylated at ITIM and ITSM motifs leading to recruitment of protein tyrosine phosphatases SHP-1 and SHP-2 and suppression of TCR signaling. CTLA-4 protein is primarily expressed on T cells, including CD8+ cytotoxic T cells, CD4+ helper T cells, and CD4+/FoxP3+ regulatory T cells. CTLA-4 protein competes with CD28 for B7.1 (CD80) and B7.2 (CD86) binding at the cell surface, resulting in downregulation of T cell activity. TIGIT is expressed at low levels on subsets of T cells and natural killer (NK) cells, and is upregulated at the protein level following activation of these cells. TIGIT marks exhausted T cells in the tumor microenvironment and during human immunodeficiency virus (HIV) infection. TIM-3 is expressed by exhausted T cells in the settings of chronic infection and cancer. Tumor-infiltrating macrophages and dendritic cells also express TIM-3. LAG3 is primarily expressed by activated CD4+ T cells, CD8+ T cells, FoxP3+ T regulatory cells (Tregs), and NK cells. 2B4 is a heterophilic cell surface receptor expressed on a variety of immune cells, including NK cells, T cells, eosinophils, mast cells, and dendritic cells. 2B4 has been shown to have both immune stimulatory and inhibitory effects on cells. Co-expression of multiple immune checkpoints help characterize the extent to which a T cell is exhausted and will respond to antigen stimulation. Therapeutic blockade of several of these immune checkpoint receptors is a promising strategy for neoplastic intervention by enabling anti-tumor immune responses (6-13).

Pathways & Proteins

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Database Links

UniProt ID: Q9BZW8 , P16410 , O95936 , P18627 , Q15116 , P36402 , Q495A1 , Q8TDQ0 , O94900 , Q96NM4

Entrez-Gene Id: 51744 , 1493 , 8320 , 3902 , 5133 , 6932 , 201633 , 84868 , 9760 , 84969

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使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

D2G4O is a trademark of Cell Signaling Technology. Inc.

D5D5R is a trademark of Cell Signaling Technology. Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

Human Exhausted T Cell Antibody Sampler Kit #81490