Human PD-L1/B7-H1 Antibody

Name: Human PD-L1/B7-H1 Antibody
Brand: R&D Systems
SKU: AF156-SP
Rating: 4.7 (3 reviews)

AF156 8 Citations 3 Reviews Datasheet / COA / SDS

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AF156
AF156-SP

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Human PD-L1/B7-H1 Antibody in Western Blot

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Citations (8)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human PD-L1/B7-H1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human PD-L1/B7-H1 in direct ELISAs and Western blots. In direct ELISAs, less than 15% cross-reactivity with recombinant mouse PD-L1/B7-H1 is observed and less than 1% cross‑reactivity with recombinant human PD-L2 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human PD-L1/B7-H1
Phe19-Thr239
Accession # Q9NZQ7

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

See below

Simple Western

50 µg/mL

See below

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561R).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human PD-L1/B7-H1 DuoSet ELISA Kit (Catalog # DY156) for convenient development of a sandwich ELISA or the Human/Cynomolgus Monkey PD-L1/B7-H1 Quantikine ELISA Kit (Catalog # DB7H10) for a complete optimized ELISA.

Immunohistochemistry

5-15 µg/mL

See below

Blockade of Receptor-ligand Interaction

In a functional ELISA, 1-5 µg/mL of this antibody will block 50% of the binding of 100 ng/mL of biotinylated Recombinant Human PD-L1/B7-H1 to immobilized Recombinant Mouse PD-1 Fc Chimera (Catalog # 1021-PD) coated at 1 µg/mL (100 µL/well). At 50 µg/mL, this antibody will block >90% of the binding.

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

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Detection of Human PD-L1/B7-H1 by Western Blot. Western blot shows lysates of human placenta tissue. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF156) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PD-L1/B7-H1 at approximately 50-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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PD-L1/B7-H1 in Human Colon and Colon Cancer Tissue. PD-L1/B7-H1 was detected in immersion fixed paraffin-embedded sections of normal human colon (left panel) and human colon cancer tissue (right panel) using Goat Anti-Human PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF156) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

ELISA

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Human PD-L1/B7-H1 ELISA Standard Curve. Recombinant Human PD-L1/B7-H1 protein was serially diluted 2-fold and captured by Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561R) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF156) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Simple Western

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Detection of Human PD-L1/B7-H1 by Simple Western^TM. Simple Western lane view shows lysates of HDLM-2 human Hodgkin's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PD-L1/B7-H1 at approximately 72 kDa (as indicated) using 50 µg/mL of Goat Anti-Human PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF156) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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

Human B7 homolog 1 (B7-H1), also called programmed cell death 1 ligand 1 (PDCD1L1) and programmed death ligand 1 (PDL1), is a member of the growing B7 family of immune proteins that provide signals for both stimulating and inhibiting T cell activation. Other family members include B7-1, B7-2, B7-H2, PDL2 and B7-H3. B7 proteins are members of the immunoglobulin (Ig) superfamily, their extracellular domains contain 2 Ig-like domains and all members have short cytoplasmic domains. Among the family members, they share about 20-25% amino acid identity. Human and mouse B7-H1 share approximately 70% amino acid sequence identity. B7-H1 has been identified as one of two ligands for programmed death-1 (PD-1), a member of the CD28 family of immunoreceptors. The B7-H1 gene encodes a 290 amino acid (aa) type I membrane precursor protein with a putative 18 aa signal peptide, a 221 aa extracellular domain, a 21 aa transmembrane region, and a 31 aa cytoplasmic domain. Human B7-H1 is constitutively expressed in several organs such as heart, skeletal muscle, placenta and lung, and in lower amounts in thymus, spleen, kidney and liver. B7-H1 expression is upregulated in a small fraction of activated T and B cells and a much larger fraction of activated monocytes. B7-H1 expression is also induced in dendritic cells and keratinocytes after IFN-gamma stimulation. Interaction of B7-H1 with PD-1 results in inhibition of TCR-mediated proliferation and cytokine production. The B7-H1:PD-1 pathway is involved in the negative regulation of some immune responses and may play an important role in the regulation of peripheral tolerance.

References

Nishimura, H. and T. Honjo (2001) Trends in Immunology 22:265.
Freeman, G.J. et al. (2000) J. Exp. Med. 192:1027.
Latchman, Y. et al. (2001) Nat. Immunol. 2:261.

Long Name

Programmed Death Ligand 1

Entrez Gene IDs

29126 (Human); 60533 (Mouse); 499342 (Rat); 574058 (Porcine); 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

Product Datasheets

Product Datasheet

COA

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Related Research Areas

Citations for Human PD-L1/B7-H1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Restoration of T Cell function in multi-drug resistant bacterial sepsis after interleukin-7, anti-PD-L1, and OX-40 administration
Authors: LK Thampy, KE Remy, AH Walton, Z Hong, K Liu, R Liu, V Yi, CD Burnham, RS Hotchkiss
PLoS ONE, 2018;13(6):e0199497.
Species: Human
Sample Types: Whole Cells
Applications: Inhibition
Immunological Properties of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells
Authors: M Idelson, R Alper, A Obolensky, N Yachimovic, J Rachmilewi, A Ejzenberg, E Beider, E Banin, B Reubinoff
Stem Cell Reports, 2018;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization
Blocking the PD-1/PD-L1 pathway in glioma: a potential new treatment strategy
Authors: S Xue, M Hu, V Iyer, J Yu
J Hematol Oncol, 2017;10(1):81.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot
Hepatitis C Virus Induces MDSCs-Like Monocytes through TLR2/PI3K/AKT/STAT3 Signaling
Authors: N Zhai, H Li, H Song, Y Yang, A Cui, T Li, J Niu, IN Crispe, L Su, Z Tu
PLoS ONE, 2017;12(1):e0170516.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
Mesenchymal Stromal Cell Secretion of Programmed Death-1 Ligands Regulates T Cell Mediated Immunosuppression
Stem Cells, 2016;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Immunoprecipitation
Soluble co-signaling molecules predict long-term graft outcome in kidney-transplanted patients.
Authors: Melendreras S, Martinez-Camblor P, Menendez A, Bravo-Mendoza C, Gonzalez-Vidal A, Coto E, Diaz-Corte C, Ruiz-Ortega M, Lopez-Larrea C, Suarez-Alvarez B
PLoS ONE, 2014;9(12):e113396.
Species: Human
Sample Types: Serum
Applications: ELISA Development
12-O-tetradecanoyl phorbol 13-acetate induces the expression of B7-DC, -H1, -H2, and -H3 in K562 cells.
Authors: Jang BC, Park YK, Choi IH, Kim SP, Hwang JB, Baek WK, Suh MH, Mun KC, Suh SI
Int. J. Oncol., 2007;31(6):1439-47.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry
Aberrant regulation of synovial T cell activation by soluble costimulatory molecules in rheumatoid arthritis.
Authors: Wan B, Nie H, Liu A, Feng G, He D, Xu R, Zhang Q, Dong C, Zhang JZ
J. Immunol., 2006;177(12):8844-50.
Species: Human
Sample Types: Serum
Applications: ELISA Development