Revision 8

#45208Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, FC-L

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

45-70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q8TDQ0

Entrez-Gene Id:

84868

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:200 - 1:800
Flow Cytometry (Live) 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #81229.

Specificity / Sensitivity

TIM-3 (D5D5R) XP® Rabbit mAb recognizes endogenous levels of total TIM-3 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the extracellular domain of human TIM-3 protein.

Background

T cell Ig- and mucin-domain-containing molecules (TIMs) are a family of transmembrane proteins expressed by various immune cells. TIM-3 is an inhibitory molecule that is induced following T cell activation (1-3 ). TIM-3 is expressed by exhausted T cells in the settings of chronic infection and cancer (4,5), and tumor-infiltrating T cells that coexpress PD-1 and TIM-3 exhibit the most severe exhausted phenotype (5). Tumor-infiltrating dendritic cells (DCs) also express TIM-3. TIM-3 expression on DCs was found to suppress innate immunity by reducing the immunogenicity of nucleic acids released by dying tumor cells (6). Research studies show that heterodimerization of TIM-3 with CEACAM-1 is critical for the inhibitory function of TIM-3, and co-blockade of TIM-3 and CEACAM-1 enhanced anti-tumor responses in a mouse model of colorectal cancer (7). In addition, blockade of TIM-3 in mouse models of autoimmunity enhanced the severity of disease (1). Finally, binding of Galectin-9 to TIM-3 expressed by Th1 cells induces T cell death (8).

  1. Monney, L. et al. (2002) Nature 415, 536-41.
  2. Sánchez-Fueyo, A. et al. (2003) Nat Immunol 4, 1093-101.
  3. Sabatos, C.A. et al. (2003) Nat Immunol 4, 1102-10.
  4. Jones, R.B. et al. (2008) J Exp Med 205, 2763-79.
  5. Sakuishi, K. et al. (2010) J Exp Med 207, 2187-94.
  6. Chiba, S. et al. (2012) Nat Immunol 13, 832-42.
  7. Huang, Y.H. et al. (2015) Nature 517, 386-90.
  8. Zhu, C. et al. (2005) Nat Immunol 6, 1245-52.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-L: Flow Cytometry (Live)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 8
#45208

TIM-3 (D5D5R) XP® Rabbit mAb

Western Blotting Image 1: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
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).
Western Blotting Image 2: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using TIM-3 (D5D5R) XP® Rabbit mAb #45208 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 3: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
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.
Immunoprecipitation Image 1: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Immunoprecipitation of TIM-3 from RPMI 8226 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TIM-3 (D5D5R) XP® Rabbit mAb, #45208. Western blot was performed using TIM-3 (D5D5R) XP® Rabbit mAb. Secondary detection was performed using #12291, Protein A (HRP Conjugate).
Immunohistochemistry Image 1: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded renal clear cell carcinoma using TIM-3 (D5D5R) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using TIM-3 (D5D5R) XP® Rabbit mAb.
Immunohistochemistry Image 2: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TIM-3 (D5D5R) XP® Rabbit mAb.
Immunohistochemistry Image 3: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TIM-3 (D5D5R) XP® Rabbit mAb. Note staining of alveolar macrophages.
Immunohistochemistry Image 4: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
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).
Immunohistochemistry Image 5: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human breast carcinoma usng TIM-3 (D5D5R) XP® rabbit mAb (yellow), PD-1 (D4W2J) XP® rabbit mAb #86163 (green), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), LAG3 (D2G4O) XP® rabbit mAb #15372 (magenta), CD8α (C8/144B) mouse mAb #70306 (orange), and Pan-keratin (C11) mouse mAb #4545 (cyan).
Immunohistochemistry Image 6: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using TIM-3 (D5D5R) XP® rabbit mAb (yellow), 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 LAG3 (D2G4O) XP® rabbit mAb #15372 (orange).
Immunohistochemistry Image 7: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using TIM-3 (D5D5R) XP® rabbit mAb (yellow), PD-1 (D4W2J) XP® rabbit mAb #86163 (green), B7-H3 (D9M2L) XP® rabbit mAb #14058 (red), B7-H4 (D1M8I) XP® rabbit mAb (cyan), LAG3 (D2G4O) XP® rabbit mAb #15372 (magenta), and VISTA (D1L2G) XP® rabbit mAb #64953 (orange).
Flow Cytometry Image 1: TIM-3 (D5D5R™) XP® Rabbit mAb Expand Image
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).