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PD-L1/CD274 Monoclonal antibody

PD-L1/CD274 Monoclonal Antibody for FC, IF, IHC, WB, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, pig, rat

Applications

WB, IP, IHC, IF, FC, CoIP, ELISA

Conjugate

Unconjugated

CloneNo.

2B11D11

Cat no : 66248-1-Ig

Synonyms

B7 H, B7 H1, B7 homolog 1, B7H1, CD274, CD274 molecule, PD L1, PDCD1 ligand 1, PDCD1L1, PDCD1LG1, PDL1, PD-L1, PD-L1/CD274, Programmed death ligand 1



Tested Applications

Positive WB detected inA375 cells, human placenta tissue, pig lung tissue, human skeletal muscle tissue, HepG2 cells, THP-1 cells, RAW 264.7 cells, A549 cells, K-562 cells, HSC-T6 cells
Positive IHC detected inhuman tonsillitis tissue, human heart tissue, human lung cancer tissue, human placenta tissue, mouse heart 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 detected inHeLa cells
Positive FC detected inMDA-MB-231 cells
Planning an IF experiment? We recommend our CoraLite® Plus 488 conjugated versions of this antibody.

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:2000-1:10000
Immunohistochemistry (IHC)IHC : 1:5000-1:20000
Immunofluorescence (IF)IF : 1:50-1:500
Flow Cytometry (FC)FC : 0.20 ug per 10^6 cells in a 100 µl suspension
Sample-dependent, check data in validation data gallery

Product Information

66248-1-Ig targets PD-L1/CD274 in WB, IP, IHC, IF, FC, CoIP, ELISA applications and shows reactivity with human, mouse, pig, rat samples.

Tested Reactivity human, mouse, pig, rat
Cited Reactivity human, rat, mouse
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Immunogen PD-L1/CD274 fusion protein Ag12443 相同性解析による交差性が予測される生物種
Full Name CD274 molecule
Calculated molecular weight 290 aa, 33 kDa
Observed molecular weight 45-50 kDa
GenBank accession numberBC074984
Gene symbol CD274
Gene ID (NCBI) 29126
RRIDAB_2756526
Conjugate Unconjugated
Form Liquid
Purification Method Protein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

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).


Protocols

Product Specific Protocols
WB protocol for PD-L1/CD274 antibody 66248-1-IgDownload protocol
IHC protocol for PD-L1/CD274 antibody 66248-1-IgDownload protocol
IF protocol for PD-L1/CD274 antibody 66248-1-IgDownload protocol
FC protocol for PD-L1/CD274 antibody 66248-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanIF

Cell Res

Targeting ATAD3A-PINK1-mitophagy axis overcomes chemoimmunotherapy resistance by redirecting PD-L1 to mitochondria

Authors - Xiao-Qing Xie
humanWB

Cell

Developing Covalent Protein Drugs via Proximity-Enabled Reactive Therapeutics.

Authors - Qingke Li
humanWB,IHC,IF

Cancer Cell

ADORA1 Inhibition Promotes Tumor Immune Evasion by Regulating the ATF3-PD-L1 Axis.

Authors - Hong Liu
human,mouseIHC

Cell Metab

NAD+ Metabolism Maintains Inducible PD-L1 Expression to Drive Tumor Immune Evasion.

Authors - Hongwei Lv
humanWB

ACS Nano

Self-Assembly of Podophyllotoxin-Loaded Lipid Bilayer Nanoparticles for Highly Effective Chemotherapy and Immunotherapy via Downregulation of Programmed Cell Death Ligand 1 Production.

Authors - Rui Wang
mouseIF

ACS Nano

Mitochondria-Targeting Polymer Micelle of Dichloroacetate Induced Pyroptosis to Enhance Osteosarcoma Immunotherapy.

Authors - Jiakang Jin

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

hala (Verified Customer) (01-27-2023)

there is 2 bands one at the specific size and the other one is higher

  • Applications: Western Blot
  • Primary Antibody Dilution: 1/4000
  • Cell Tissue Type: hepG2
PD-L1/CD274 Antibody Western Blot validation (1/4000 dilution) in hepG2 (Cat no:66248-1-Ig)
FH

Emma (Verified Customer) (11-29-2021)

Used for IF on FFPE prostate tissue at 1:50 with Tris-EDTA antigen retrieval (pH 9.0).

  • Applications: Immunofluorescence
  • Primary Antibody Dilution: 1:50
  • Cell Tissue Type: Prostate
PD-L1/CD274 Antibody Immunofluorescence validation (1:50 dilution) in Prostate (Cat no:66248-1-Ig)
FH

Marina (Verified Customer) (06-14-2021)

HCC1937 breast cancer cells were fixed with 4% PFA, permeabilized and stained overnight at 4ºC with anti-PD-L1 Ig diluted 1:300. Nuclei were stained with DAPI.

  • Applications: Immunofluorescence
  • Primary Antibody Dilution: 1:300
  • Cell Tissue Type: Immortalized human breast cancer cells
PD-L1/CD274 Antibody Immunofluorescence validation (1:300 dilution) in Immortalized human breast cancer cells (Cat no:66248-1-Ig)