PD-L1/CD274 Monoclonal antibody, PBS Only

• Featured Product
• KD/KO Validated

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

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, pig, rat

Applications

WB, IF, FC, IHC, ELISA

Conjugate

Unconjugated

CloneNo.

2B11D11

Cat no : 66248-1-PBS

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

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Validation Data Gallery

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  WB analysis using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  WB analysis of human placenta using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  WB analysis of human skeletal muscle using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  WB analysis of RAW 264.7 using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  WB analysis of human placenta using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  WB analysis of HepG2 using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  WB analysis of THP-1 using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  WB analysis of HepG2 cells using 66248-1-Ig (same clone as 66248-1-PBS)

  WB result of PD-L1 antibody (66248-1-Ig, 1:1000 and 1:2000) with si-Control and si-PDL1 transfected HepG2 cells. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  WB analysis of pig lung using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  IHC staining of human tonsillitis using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human tonsillitis using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  IHC staining of human heart using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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  IHC staining of human heart using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of mouse heart using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of mouse heart using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human lung cancer using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human placenta using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human placenta using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human lung cancer using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  IF Staining of HeLa using 66248-1-Ig (same clone as 66248-1-PBS)

  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). This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

• 

  FC experiment of MDA-MB-231 using 66248-1-Ig (same clone as 66248-1-PBS)

  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. This data was developed using the same antibody clone with 66248-1-PBS in a different storage buffer formulation.

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Recommended dilution

Application	Dilution
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.

Product Information

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

• Tested Reactivity: human, mouse, pig, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: PD-L1/CD274 fusion protein Ag12443
• Full Name: CD274 molecule
• Calculated Molecular Weight: 290 aa, 33 kDa
• Observed Molecular Weight: 45-50 kDa
• GenBank Accession Number: BC074984
• Gene Symbol: CD274
• Gene ID (NCBI): 29126
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS only
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

Programmed cell death ligand 1 (PD-L1, CD274, or B7-H1), is the first member of B7 family to be discovered. B7 family molecules are type I transmembrane proteins belonging to the immunoglobulin superfamily. In concert with their CD28 family receptors, the B7s are key regulators of the adaptive immune response. PD-L1 is suggested as a negative regulator of T and B cell, and plays important role in mediating tolerance of lymphocytes to self-antigens. It is also involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PD-1-independent manner. PD-L1 is a 290 aa transmembrane protein with a calculated molecular weight of 33 kDa, it is predicted to be 27-30 kDa after signal peptide cleavage (PMID: 25609200; 17076679). The apparent molecular weight has also been reported as 45-70 kDa, major glycosylated form of 45-50 kDa and multiple post-translational modifications form of 65-70 kDa (PMID: 18760278; 16493058).