Human PD-L1 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB1562-100
MAB1562-SP
Detection  of human PD-L1/B7-H1 by Western Blot.
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Human PD-L1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PD-L1/B7-H1 in direct ELISAs.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2340D
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Mouse myeloma cell line, NS0-derived human PD-L1/B7-H1 protein
Phe19-Thr239
Accession # Q9NZQ7
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Blockade of Receptor-ligand Interaction
In a functional ELISA binding assay, 0.2-1.2 µg/mL of thisantibody will block 50% of the binding of 1 µg/mL of Recombinant HumanPD-L1/B7-H1 (Catalog # 156-B) to immobilized Recombinant Human PD-1 Fc chimera (Catalog # 1086-PD) coated at 100 ng/mL (100 µL/well). At 2-3 µg/mL, this antibodywill block >90% of the binding.
 
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection  of human PD-L1/B7-H1 by Western Blot. View Larger

Detection of human PD-L1/B7-H1 by Western Blot. Western blot shows lysates of human lung tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1562) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for PD-L1/B7-H1 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry Detection of PD-L1/B7-H1 in HEK293 Human Cell Line Transfected with Human PD-L1/B7-H1  and eGFP by Flow Cytometry. View Larger

Detection of PD-L1/B7-H1 in HEK293 Human Cell Line Transfected with Human PD-L1/B7-H1 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human PD-L1/B7-H1 or (B) irrelevant protein and eGFP was stained with Rabbit Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1562) followed by APC-conjugated Goat anti-Rabbit IgG Secondary Antibody (Catalog # F0111). Quadrant markers were set based on Rabbit IgG control antibody staining (Catalog # MAB1050). View our protocol for Staining Membrane-associated Proteins.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PD-L1

B7-H1, also known as PD-L1 and CD274, is an approximately 65 kDa transmembrane glycoprotein in the B7 family of immune regulatory molecules (1). Mature human B7-H1 consists of a 220 amino acid (aa) extracellular domain (ECD) with two immunoglobulin-like domains, a 21 aa transmembrane segment, and a 31 aa cytoplasmic domain (2). Within the ECD, human B7-H1 shares 73% and 74% aa sequence identity with mouse and rat B7-H1, respectively. Alternative splicing generates additional isoforms that either lack the first Ig-like domain or are truncated within the second Ig-like domain (3). B7-H1 is expressed on
inflammatory-activated immune cells including macrophages, T cells, and B cells (4-7), keratinocytes (8, 9), enothelial and intestinal epithelial cells (8, 10), as well as a variety of carcinomas and melanoma (11, 12). B7-H1 binds to T cell B7-1/CD80 and PD-1 (7, 8, 12-15). It suppresses T cell activation and proliferation (5, 8, 14, 16) and induces the apoptosis of activated T cells (11). It plays a role in the development of immune tolerance by promoting T cell anergy (7, 14) and enhancing regulatory T cell development (16). B7-H1 favors the development of anti-inflammatory IL-10 and IL-22 producing dendritic cells (5, 10) and inhibits the development of Th17 cells (16). In cancer, B7-H1 provides resistance to T cell mediated lysis, enhances EMT, and enhances the tumorigenic function of Th22 cells (6, 9, 12, 15).

References
  1. Ceeraz, S. et al. (2013) Trends Immunol. 34:556.
  2. Dong, H. et al. (1999) Nat. Med. 5:1365.
  3. Frigola, X. et al. (2011) Clin. Cancer Res. 17:1915.
  4. Tamura, H. et al. (2001) Blood 97:1809.
  5. Chen, L. et al. (2007) J. Immunol. 178:6634.
  6. Kuang, D.-M. et al. (2014) J. Clin. Invest. 124:4657.
  7. Tsushima, F. et al. (2007) Blood 110:180.
  8. Mazanet, M.M. and C.C.W. Hughes (2002) J. Immunol. 169:3581.
  9. Cao, Y. et al. (2010) Cancer Res. 71:1235.
  10. Scandiuzzi, L. et al. (2014) Cell Rep. 6:625.
  11. Dong, H. et al. (2002) Nat. Med. 8:793.
  12. Azuma, T. et al. (2008) Blood 111:3635.
  13. Butte, M.J. et al. (2008) Mol. Immunol. 45:3567.
  14. Park, J.-J. et al. (2010) Blood 116:1291.
  15. Ritprajak, P. et al. (2010) J. Immunol. 184:4918.
  16. Herold, M. et al. (2015) J. Immunol. 195:3584.
Long Name
Programmed Death Ligand 1
Entrez Gene IDs
29126 (Human); 60533 (Mouse); 499342 (Rat); 484186 (Canine); 102145573 (Cynomolgus Monkey)
Alternate Names
B7-H; B7H1; B7-H1; B7H1PDCD1L1; CD274 antigenMGC142294; CD274 molecule; CD274; PDCD1L1; PDCD1LG1; PDCD1LG1MGC142296; PDL1; PD-L1; PD-L1B7 homolog 1; PDL1PDCD1 ligand 1; programmed cell death 1 ligand 1; Programmed death ligand 1

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