PDL1 Antibody [6H10]

PDL1 Antibody [6H10] | RF16035 | ProSci

Host: Mouse

Reactivity: Human, Mouse

Homology: Predicted species reactivity based on immunogen sequence: Rat: (77%) , Mouse: (71%)

Immunogen: Anti-PD-L1 antibody (RF16035) was raised against the extracellular domain of human PD-L1.

Research Area: Immunology

Tested Application: E, WB, IHC-P, ICC, IF

Application: WB: 0.25 - 4 μg/mL; IF: 2 μg/mL; IHC-P: 5 μg/mL; ICC: 1 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry, Immunocytochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.

Specificiy: PD-L1 Antibody has no cross-reactivity to PD-L2.

Positive Control 1: Cat. No. 1201 - HeLa Cell Lysate

Positive Control 2: Cat. No. 1224 - Daudi Cell Lysate

Positive Control 3: Cat. No. 1340 - Human Stomach Carcinoma Tissue Lysate

Positive Control 4: Cat. No. 1315 - Human Tonsil Tissue

Positive Control 5: N/A

Positive Control 6: N/A

Molecular Weight: Predicted: 33kD
Observed: 37kD

Validation:

Independent Antibody Validation (Figure 2) shows similar PD-L1 expression profile in both human and mouse cell lines detected by two independent anti-PD-L1 antibodies that recognize different epitopes, RF16035 against the center of human PD-L1 and RF16035 against the extracellular domain.  PD-L1 proteins are detected in most of the cell lines, but not in A549 and THP-1 cells by the two independent antibodies. 

siRNA Knockdown Validation (Figure 3) : Anti-PD-L1 antibody (RF16035) specificity was further verified by PD-L1 specific siRNA knockdown. PD-L1 signal in HeLa cells transfected with PD-L1 siRNAs was much weaker in comparison with that in HeLa cells transfected with control siRNAs.

Overexpression Validation (Figure 1, 5, 10) : Anti-PD-L1 antibodies (RF16035) can detect the overexpression of PD-L1 protein in 293 cells transfected with PD-L1.

Isoform: Human PD-L1 has 3 isoforms, including isoform 1 (290aa, 33.3kD) , isoform 2 (176aa, 20.2kD) , and isoform 3 (178aa, 20.5kD) . This antibody detects human isoform 1&3, mouse and rat PD-L1 (290aa, 33kD for both of them) .

Purification: PD-L1 Antibody is supplied as protein A purified IgG1.

Clonality: Monoclonal

Clone: 6H10

Isotype: IgG1

Conjugate: Unconjugated

Physical State: Liquid

Buffer: PD-L1 Antibody is supplied in PBS containing 0.02% sodium azide and 50% glycerol.

Concentration: 1 mg/mL

Storage Condition: PD-L1 antibody can be stored at 4˚C for three months and -20˚C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Alternate Name: PD-L1 Antibody: Programmed cell death 1 ligand-1, programmed death ligand 1, PDL1, PDL-1, B7-H1

User Note: Optimal dilutions for each application to be determined by the researcher.

BACKGROUND: PD-L1 plays a critical role in induction and maintenance of immune tolerance to self. As a ligand for the inhibitory receptor PDCD1/CD279, PD-L1 modulates the activation threshold of T-cells and limits T-cell effector response (1) . The PDCD1/CD279-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and facilitate tumor survival (2, 3) . Through a yet unknown activating receptor, it may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (4) .