Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry, ELISA Detection, ELISA Development (Capture)

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # 130021

View all PD-L1/B7-H1 Antibodies »

Product Specifications

Immunogen

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

Specificity

Detects human PD-L1/B7-H1 in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human (rh) B7-1, -2, -H2, -H3, -H3b,-H4, rhPD-L2, recombinant mouse B7-H1, recombinant rat (rr) B7-1, or rrB7-2 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for Human PD-L1/B7-H1 Antibody

Detection of PD-L1/B7-H1 in Jurkat Human Cell Line by Flow Cytometry.

Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B).View our protocol for Staining Membrane-associated Proteins.

Detection of PD-L1/B7-H1 in MDA-MB-231 Human Cell Line by Flow Cytometry.

MDA-MB-231 human breast adenocarcinoma cell line was stained with Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). Adherent cells were prepared by either manual scraping or with TrypLE Express treatment with similar results. View our protocol for Staining Membrane-associated Proteins.

PD-L1/B7-H1 in Human Colon Cancer.

PD-L1/B7-H1 was detected in formalin fixed paraffin-embedded sections of human colon cancer using Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was observed in the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of PD-L1/B7-H1 in Human Colon Cancer.

Formalin-fixed paraffin-embedded tissue sections of human colon cancer were probed for PDL1 mRNA (ACD RNAScope Probe, catalog # 600861; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using mouse anti-human PDL1 monoclonal antibody (R&D Systems catalog # MAB1561) at 5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes.

Detection of PD-L1/B7-H1 by Western Blot

Chemotherapeutics induce PD-L1 expression in NPC cells and PD-1 expression in NK cells via upregulation of NF-kappa B. NPC cells a or NK cells c were incubated with the NF-kappa B inhibitor BMS-345541 for 1 h before incubation with chemotherapeutics. NPC cells were transfected with NF-kappa B siRNA b or NK cells with NF-kappa B siRNA d for 16 h and then incubated with chemotherapeutics. Expression of NF-kappa B, PD-L1 and PD-1 was analyzed by immunoblot Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32737537), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PD-L1/B7-H1 by Western Blot

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Applications for Human PD-L1/B7-H1 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Dual RNAscope ISH-IHC Compatible

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Jurkat human acute T cell leukemia cell line or MDA-MB-231 human breast adenocarcinoma cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon

Reviewed Applications

Read 5 reviews rated 4.8 using MAB1561 in the following applications:

Flow Cytometry (1 Review)
Immunocytochemistry/Immunofluorescence (1 Review)
Immunohistochemistry (1 Review)
SPR (1 Review)
SPR ASSAY (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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

Human B7 homolog 1 (B7-H1), also called programmed death ligand 1 (PD-L1) and programmed cell death 1 ligand 1 (PDCD1L1), 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, there is 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 291 amino acid (aa) type I membrane precursor protein with a putative 18 aa signal peptide, a 220 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 Immunol. 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

Alternate Names

B7-H1, B7H1, CD274, PDCD1L1, PDCD1LG1, PDL1

Entrez Gene IDs

29126 (Human); 60533 (Mouse); 499342 (Rat); 574058 (Porcine); 484186 (Canine); 102145573 (Cynomolgus Monkey)

Gene Symbol

CD274

UniProt

Q9NZQ7

Additional PD-L1/B7-H1 Products

Product Documents for Human PD-L1/B7-H1 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human PD-L1/B7-H1 Antibody

For research use only

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