PD-L1/CD274 (C-terminal) Polyclonal antibody

• Featured Product
• KD/KO Validated

PD-L1/CD274 (C-terminal) Polyclonal Antibody for WB, IHC, IF-P, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF-P, IP, ChIP, ELISA

Conjugate

Unconjugated

Publications

91

Cat no : 28076-1-AP

Synonyms

CD274, PD-L1, PDCD1 ligand 1, PD L1, hPD-L1

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Validation Data Gallery

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  WB analysis using 28076-1-AP

  Untreated and PNGase F-treated lysates of human placenta were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours. PNGase F was obtained from Atagenix (cat.NO. ata808).

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  WB analysis using 28076-1-AP

  Lysates of untreated and IFN gamma treated A549 cells were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse heart using 28076-1-AP

  mouse heart tissue were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 28076-1-AP

  MCF-7 cells (PD-L1-), MDA-MB-231 cells (PD-L1+), and human placenta tissue were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  WB analysis of THP-1 using 28076-1-AP

  WB result of PD-L1/CD274 (C-terminal) antibody (28076-1-AP; 1:300; incubated at room temperature for 1.5 hours) with sh-Control and sh-PD-L1/CD274 (C-terminal) transfected THP-1 cells.

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  WB analysis of human placenta using 28076-1-AP

  human placenta tissue were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat heart using 28076-1-AP

  rat heart tissue were subjected to SDS PAGE followed by western blot with 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  IP experiment of MDA-MB-231 using 28076-1-AP

  IP result of anti-PD-L1/CD274 (C-terminal) (IP:28076-1-AP, 4ug; Detection:28076-1-AP 1:3000) with MDA-MB-231 cells lysate 920 ug.

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  IHC staining of human tonsillitis using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human cervical cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human cervical cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:400 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:800 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 28076-1-AP (PD-L1/CD274 antibody) at dilution of 1:800 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 28076-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human tonsillitis using 28076-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using PD-L1/CD274 (C-terminal) antibody (28076-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CoraLite®594 CD3 antibody (CL594-60181, Clone: 3F3A1, red).

• 

  IF Staining of human placenta using 28076-1-AP

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded human placenta tissue using PD-L1/CD274 (C-terminal) antibody (28076-1-AP) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human tonsillitis using 28076-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:50 and CoraLite488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of human tonsillitis using 28076-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using 28076-1-AP (PD-L1/CD274 (C-terminal) antibody) at dilution of 1:50 and CoraLite488-Conjugated Goat Anti-Rabbit IgG(H+L).

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Tested Applications

• Positive WB detected in: IFN gamma treated A549 cells, mouse heart tissue, rat heart tissue, MDA-MB-231 cells, human placenta tissue, THP-1 cells
• Positive IP detected in: MDA-MB-231 cells
• Positive IHC detected in: human tonsillitis tissue, human placenta tissue, human breast cancer tissue, human lung cancer tissue, human cervical cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human tonsillitis tissue, human placenta tissue

Recommended dilution

• Western Blot (WB): WB : 1:300-1:1000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 59 publications below
• IHC: See 37 publications below
• IF: See 18 publications below
• IP: See 1 publications below
• ELISA: See 1 publications below
• ChIP: See 1 publications below

Product Information

28076-1-AP targets PD-L1/CD274 (C-terminal) in WB, IHC, IF-P, IP, ChIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: PD-L1/CD274 (C-terminal) fusion protein Ag27557
• Full Name: CD274 molecule
• Calculated Molecular Weight: 290 aa, 33 kDa
• Observed Molecular Weight: 45-50 kDa
• GenBank Accession Number: BC074984
• Gene Symbol: PD-L1
• Gene ID (NCBI): 29126
• RRID: AB_2881052
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

PD-L1, also known as CD274 or B7H1, stands for programmed cell death ligand 1. It is a type I transmembrane protein that is thought to repress immune responses by binding to its receptor (PD1), thus inhibiting T-cell activation, proliferation, and cytokine production. It contains V-like and C-like immunoglobulin domains. PD-L1 expression is regulated by various cytokines, such as TNF-α or LPS (ISSN: 1848-7718). Increased expression of this protein in certain types of cancers, e.g., renal cell carcinoma or colon cancer, correlates with poor prognosis. 

What is the molecular weight of PD-L1? 

Depending on the isoform, the calculated molecular weight of the protein varies between 20 and 33 kDa (176-290 aa). 

What are the isoforms of PD-L1? 

According to NCBI, three different isoforms have been identified. There are significant differences in the untranslated and protein coding regions. 

What is the subcellular localization and tissue specificity of PD-L1? 

It is predicted to localize in the plasma membrane of various cell types, with a particularly high expression in placental trophoblast and subsets of immune cells. High levels of PD-L1 protein have also been detected in lung and colon tissues. 

What is the function of PD-L1 in immune responses? 

PD-L1 is critical for the induction and maintenance of immune self-tolerance during infection or inflammation in normal tissues. The interaction of PD-L1 and its receptors is responsible for preventing auto-immune phenotypes and balancing the overall immune response in situations such as pregnancy or tissue allografts. The interaction between PD-L1 and PD-1 or B7.1 starts an inhibitory signaling cascade, which results in the decreased proliferation of antigen-specific T-cells and increased survival of regulatory T-cells (PMID: 15240681). 

How can PD-L1's implication in cancer be used as a drug target? 

In certain tumors, high expression of PD-L1 serves as a stop-sign to inhibit the recognition of cancer cells by T-cells (PMID: 23087408). The interaction between PD-L1 and its receptors (PD1 and B7.1) is a mechanism for the tumor to evade the host immune response (PMID: 29357948). Several mAbs have been developed to target that interaction and thus prevent the inactivation of cytotoxic T-cells by the tumor (PMIDs: 23890059, 18173375).

Publications

Species	Application	Title
mouse	IHC	ACS Cent Sci; Allosteric Regulation of IGF2BP1 as a Novel Strategy for the Activation of Tumor Immune Microenvironment; Authors - Yang Liu
human	IF	Cell Rep Med; Benzosceptrin C induces lysosomal degradation of PD-L1 and promotes antitumor immunity by targeting DHHC3; Authors - Qun Wang
human,mouse	WB,IHC	Sci Adv; Inhibition of ACLY overcomes cancer immunotherapy resistance via polyunsaturated fatty acids peroxidation and cGAS-STING activation; Authors - Wei Xiang
mouse	WB	Sci Adv; Promoting the activation of T cells with glycopolymer-modified dendritic cells by enhancing cell interactions.; Authors - Liyin Yu
human	WB	Biosens Bioelectron; Aptamer-bivalent-cholesterol-mediated proximity entropy-driven exosomal protein reporter for tumor diagnosis; Authors - Zhichao Fan
human,mouse	WB,IHC	J Exp Clin Cancer Res; Anoikis resistance and immune escape mediated by Epstein-Barr virus-encoded latent membrane protein 1-induced stabilization of PGC-1α promotes invasion and metastasis of nasopharyngeal carcinoma; Authors - Chaoliang Liao