Revision 9
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, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

52-65

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q15116

Entrez-Gene Id:

5133

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
IHC Leica Bond 1:100 - 1:400
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:200

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 #63815.

Specificity / Sensitivity

PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb recognizes endogenous levels of total PD-1 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala274 of human PD-1 protein.

Background

The programmed cell death 1 protein (PD-1, PDCD1, CD279) is a member of the CD28 family of immunoreceptors that regulate T cell activation and immune responses (1-3). The PD-1 protein contains an extracellular Ig V domain, a transmembrane domain, and a cytoplasmic tail that includes an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). PD-1 is activated by the cell surface ligands PD-L1 and PD-L2 (4). Upon activation, PD-1 ITIM and ITSM phosphorylation leads to the recruitment of the protein tyrosine phosphatases SHP-1 and SHP-2, which suppress TCR signaling (5-7). In addition to activated T cells, PD-1 is expressed in activated B cells and monocytes, although its function in these cell types has not been fully characterized (8). The PD-1 pathway plays an important role in immune tolerance (3); however, research studies show that cancer cells often adopt this pathway to escape immune surveillance (9). Consequently, blockade of PD-1 and its ligands is proving to be a sound strategy for neoplastic intervention (10).

  1. Ishida, Y. et al. (1992) EMBO J 11, 3887-95.
  2. Shinohara, T. et al. (1994) Genomics 23, 704-6.
  3. Nishimura, H. et al. (1999) Immunity 11, 141-51.
  4. Freeman, G.J. et al. (2000) J Exp Med 192, 1027-34.
  5. Yokosuka, T. et al. (2012) J Exp Med 209, 1201-17.
  6. Sheppard, K.A. et al. (2004) FEBS Lett 574, 37-41.
  7. Chemnitz, J.M. et al. (2004) J Immunol 173, 945-54.
  8. Thibult, M.L. et al. (2013) Int Immunol 25, 129-37.
  9. Dong, H. et al. (2002) Nat Med 8, 793-800.
  10. Topalian, S.L. et al. (2012) Curr Opin Immunol 24, 207-12.

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) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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 9
#86163

PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb

Western Blotting Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from human CD4+ T cells, MOLT-4, and Jurkat cells using PD-1 (Intracellular Domain) (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).
Immunoprecipitation Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
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 (Intracellular Domain) (D4W2J) XP® Rabbit mAb. Western blot analysis was performed using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb.
Immunohistochemistry Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human infiltrating papillary carcinoma of the breast using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb.
Immunohistochemistry Image 2: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb.
Immunohistochemistry Image 3: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 293 cell pellets, control (left) or PD-1 transfected (right), using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb.
Immunohistochemistry Image 4: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human breast carcinoma usng PD-1 (Intracellular Domain) (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).
Immunohistochemistry Image 5: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using PD-1 (Intracellular Domain) (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).
Immunofluorescence Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of MOLT-4 cells (left, positive) or HeLa cells (right, negative) using PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb (green), Cytochrome c (6H2.B4) Mouse mAb #12963 (red), and DAPI #4083 (blue).
Flow Cytometry Image 1: PD-1 (Intracellular Domain) (D4W2J) XP® Rabbit mAb Expand Image
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 (Intracellular Domain) (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.