Anticorps Monoclonal anti-PD-L1/CD274

• Phare
• Validé par KD/KO

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

Hôte / Isotype

Mouse / IgG1

Réactivité testée

Humain, porc, rat, souris

Applications

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

Conjugaison

Non conjugué

Publications

239

CloneNo.

2B11D11

Avis (3)

N° de cat : 66248-1-Ig

Synonymes

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

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Galerie de données de validation

• 

  WB analysis using 66248-1-Ig

  Untreated and IFN gamma treated A375 cells and A549 cells were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human placenta using 66248-1-Ig

  human placenta tissue were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 cells using 66248-1-Ig

  WB result of PD-L1 antibody (66248-1-Ig, 1:1000 and 1:2000) with si-Control and si-PDL1 transfected HepG2 cells.

• 

  WB analysis of human placenta using 66248-1-Ig

  human placenta tissue were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of pig lung using 66248-1-Ig

  pig lung tissue were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human skeletal muscle using 66248-1-Ig

  human skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of RAW 264.7 using 66248-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 66248-1-Ig

  HepG2, K-562, HSC-T6, and Raw 264.7 cells were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of THP-1 using 66248-1-Ig

  Untreated and PNGase F-treated lysates of THP-1 cells and HepG2 cells were subjected to SDS PAGE followed by western blot with 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. PNGase F was obtained from Atagenix (cat.NO. ata808).

• 

  IHC staining of human tonsillitis using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:10000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:10000 (under 4x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:20000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). Slide was treated with PNGase F before staining.

• 

  IHC staining of human placenta using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:20000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). Slide was treated with PNGase F before staining.

• 

  IHC staining of mouse heart using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse heart using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:20000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). Slide was treated with PNGase F before staining.

• 

  IHC staining of human lung cancer using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66248-1-Ig (PD-L1/CD274 antibody) at dilution of 1:20000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). Slide was treated with PNGase F before staining.

• 

  IHC staining of human heart using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 66248-1-Ig (PD-L1/CD274 Antibody) at dilution of 1:500 (under 10x lens).

• 

  IHC staining of human heart using 66248-1-Ig

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 66248-1-Ig (PD-L1/CD274 Antibody) at dilution of 1:500 (under 40x lens).

• 

  IF Staining of HeLa using 66248-1-Ig

  Immunofluorescent analysis of (10% Formaldehyde ) fixed HeLa cells using 66248-1-Ig(PD-L1/CD274 antibody) at dilution of 1:300 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of MDA-MB-231 using 66248-1-Ig

  1X10^6 MDA-MB-231 cells were stained with 0.2 ug Anti-Human PD-L1/CD274 (66248-1-Ig, Clone: 2B11D11) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (green), or stained with 0.2 ug mosue IgG1 isotype control antibody and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (black). Cells were fixed with 90% MeOH.

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Applications testées

• Résultats positifs en WB: cellules A375, cellules A549, cellules HepG2, cellules HSC-T6, cellules K-562, cellules RAW 264.7, cellules THP-1, tissu de muscle squelettique humain, tissu placentaire humain, tissu pulmonaire de porc
• Résultats positifs en IHC: tissu d'amygdalite humain, tissu cardiaque de souris, tissu cardiaque humain, tissu de cancer du poumon humain, tissu placentaire humain; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en IF: cellules HeLa,
• Résultats positifs en cytométrie: cellules MDA-MB-231

Dilution recommandée

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochimie (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

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

Applications publiées

• KD/KO: See 5 publications below
• WB: See 153 publications below
• IHC: See 105 publications below
• IF: See 54 publications below
• IP: See 2 publications below
• FC: See 3 publications below
• CoIP: See 3 publications below

Informations sur le produit

66248-1-Ig cible PD-L1/CD274 dans les applications de WB, IP, IHC, IF, FC, CoIP, ELISA et montre une réactivité avec des échantillons Humain, porc, rat, souris

• Réactivité: Humain, porc, rat, souris
• Réactivité citée: rat, Humain, souris
• Hôte / Isotype: Mouse / IgG1
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: PD-L1/CD274 Protéine recombinante Ag12443
• Nom complet: CD274 molecule
• Masse moléculaire calculée: 290 aa, 33 kDa
• Poids moléculaire observé: 45-50 kDa
• Numéro d’acquisition GenBank: BC074984
• Symbole du gène: CD274
• Identification du gène (NCBI): 29126
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine A
• Tampon de stockage: PBS avec azoture de sodium à 0,02 % et glycérol à 50 % pH 7,3
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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
human	IF	Cell Res; Targeting ATAD3A-PINK1-mitophagy axis overcomes chemoimmunotherapy resistance by redirecting PD-L1 to mitochondria; Authors - Xiao-Qing Xie
human	WB	Cell; Developing Covalent Protein Drugs via Proximity-Enabled Reactive Therapeutics.; Authors - Qingke Li
human	WB,IHC,IF	Cancer Cell; ADORA1 Inhibition Promotes Tumor Immune Evasion by Regulating the ATF3-PD-L1 Axis.; Authors - Hong Liu
human,mouse	IHC	Cell Metab; NAD+ Metabolism Maintains Inducible PD-L1 Expression to Drive Tumor Immune Evasion.; Authors - Hongwei Lv
human	WB	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
mouse	IF	ACS Nano; Mitochondria-Targeting Polymer Micelle of Dichloroacetate Induced Pyroptosis to Enhance Osteosarcoma Immunotherapy.; Authors - Jiakang Jin

Avis

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

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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)

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