PDL1 Antibody [6H10] Cat. No.: RF16035

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psi-iconSpecifications
HOST SPECIES:Mouse
SPECIES 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.
TESTED APPLICATIONS: ELISA, ICC, IF, IHC-P, WB
APPLICATIONS: 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.
SPECIFICITY: PD-L1 Antibody has no cross-reactivity to PD-L2.
POSITIVE CONTROL:1) Cat. No. 1201 - HeLa Cell Lysate
 2) Cat. No. 1224 - Daudi Cell Lysate
 3) Cat. No. 1340 - Human Stomach Carcinoma Tissue Lysate
 4) Cat. No. 1315 - Human Tonsil Tissue
PREDICTED MOLECULAR WEIGHT: Predicted: 33kD

Observed: 37kD
psi-iconAdvanced Validation
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.

ISOFORMS: 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).

psi-iconProperties
PURIFICATION:PD-L1 Antibody is supplied as protein A purified IgG1.
CLONALITY:Monoclonal
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 CONDITIONS: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.

psi-iconAdditional Info
OFFICIAL SYMBOL:CD274
ALTERNATE NAMES:PD-L1 Antibody: Programmed cell death 1 ligand-1, programmed death ligand 1, PDL1, PDL-1, B7-H1
ACCESSION NO.:NP_054862
PROTEIN GI NO.:7661534
GENE ID:29126
USER NOTE:Optimal dilutions for each application to be determined by the researcher.
psi-iconBackground and References
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).
REFERENCES:1) Freeman et al. Exp. Med. 2000; 192:1027-34.
2) Burr et al. Nature 2017; 549:101-5.
3) Mezzadra et al. Nature 2017; 549:106-10.
4) Dong et al. Nat. Med. 1999 5:1365-9.

ANTIBODIES FOR RESEARCH USE ONLY.

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