Revision 6
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-F, IF-IC, FC-FP

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

40-75

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q02242

Entrez-Gene Id:

18566

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Frozen) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #55789.

Specificity / Sensitivity

PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb recognizes endogenous levels of total PD-1 protein.

Species Reactivity:

Mouse

Species predicted to react based on 100% sequence homology

Rat, Hamster

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala242 of mouse 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-F: Immunofluorescence (Frozen) 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.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 6
#84651

PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb

Western Blotting Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from EL4, CTLL-2, and NIH/3T3 using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunoprecipitation of PD-1 protein from EL4 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) (D7D5W) XP® Rabbit mAb. Western blot analysis was performed using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse LL/2 syngeneic tumor using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb performed on the Leica® Bond Rx.
Immunohistochemistry Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 mammary tumor using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 2: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 3: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse colon using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 4: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse lung using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 5: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse ovary using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb.
Immunohistochemistry Image 6: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded mouse LL2 syngeneic tumor tissue using PD-1 (Intracellular Domain) (D7D5W) XP® rabbit mAb (green), F4/80 (D2S9R) XP® rabbit mAb #70076 (orange), PD-L1 (D5V3B) rabbit mAb #64988 (yellow), CD3ε (D4V8L) rabbit mAb #99940 (red), CD8α (D4W2Z) XP® rabbit mAb #98941 (magenta) and pan keratin (C11) mouse mAb #4279 (cyan).
Immunofluorescence Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse spleen using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb has been validated on freshly prepared frozen tissue sections that were fixed for 15 minutes in 4% formaldehyde. Over-fixation diminishes the performance of this antibody.
Immunofluorescence Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of EL4 cells (positive, left) and NIH/3T3 cells (negative, right) using PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: PD-1 (Intracellular Domain) (D7D5W) XP® Rabbit mAb Expand Image
Flow cytometric analysis of CTLL-2 cells (blue) and EL4 cells (green) using PD-1 (Intracellular Domain) (D7D5W) 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.