PD-L1 Antibody Cat. No.: 4059

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psi-iconSpecifications
HOST SPECIES:Rabbit
SPECIES REACTIVITY: Human, Mouse, Rat
HOMOLOGY: Predicted species reactivity based on immunogen sequence: Rat: (77%), Mouse: (71%)
IMMUNOGEN: Anti-PD-L1 antibody (4059) was raised against a peptide corresponding to 17 amino acids near the center of human PD-L1 isoform 1.

The immunogen is located within amino acids 60 - 110 of PD-L1.
TESTED APPLICATIONS: ELISA, Flow, IF, IHC-P, WB
APPLICATIONS: WB: 0.5 - 2 μg/mL; IHC-P: 2.5 μg/mL; IF: 20 μg/mL; Flow Cyt: 0.5 μg/mL.

Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human and rat samples; Immunofluorescence in human and rat samples and Flow Cytometry in mouse 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. 1204 - K562 Cell Lysate
 4) Cat. No. 10-501 - Human Heart Tissue Slide
 5) Cat. No. 1301 - Human Heart Tissue Lysate
PREDICTED MOLECULAR WEIGHT: Predicted: 33 kDa

Observed: 37 kDa
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, 4059 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 (4059) 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 4): Anti-PD-L1 antibodies (4059) 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 affinity chromatography purified via peptide column.
CLONALITY:Polyclonal
ISOTYPE:IgG
CONJUGATE:Unconjugated
PHYSICAL STATE:Liquid
BUFFER:PD-L1 Antibody is supplied in PBS containing 0.02% sodium azide.
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: B7-H, B7H1, PDL1, PD-L1, PDCD1L1, PDCD1LG1, Programmed cell death 1 ligand 1, B7 homolog 1
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.
psi-iconCITATIONS
CITATIONS: 1)Dhar et al. Evaluation of PD-L1 expression on vortex-isolated circulating tumor cells in metastatic lung cancer. Sci Rep. 2018;8(1):2592. PMID: 29416054
2)Gadiot et al. Overall survival and PD-L1 expression in metastasized malignant melanoma. Cancer.2011;117(10):2192-201. PMID: 21523733
3)Angell et al. BRAF V600E in papillary thyroid carcinoma is associated with increased programmed death ligand 1 expression and suppressive immune cell infiltration. Thyroid. 2014; 24(9):1385-93PMID: 24955518
4)Ilie et al. Comparative study of the PD-L1 status between surgically resected specimens and matched biopsies of NSCLC patients reveal major discordances: a potential issue for anti-PD-L1 therapeutic strategies. Ann Oncol. 2016;27(1):147-53.PMID: 26483045
5)Heymann et al. PD-L1 expression in non-small cell lung carcinoma: Comparison among cytology, small biopsy, and surgical resection specimens. Cancer Cytopathol. 2017;125(12):896-907. PMID: 29024471
6)Hamanishi et al. Safety and Antitumor Activity of Anti-PD-1 Antibody, Nivolumab, in Patients With Platinum-Resistant Ovarian Cancer. J Clin Oncol. 2015; 33(34):4015-22. PMID: 26351349
7)Taube et al. Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape. Sci Transl Med. 2012;4(127):127ra37. PMID: 22461641
8)Munari et al. PD-L1 Expression Heterogeneity in Non-Small Cell Lung Cancer: Defining Criteria for Harmonization between Biopsy Specimens and Whole Sections. J Thorac Oncol. 2018;13(8): 1113-1120. PMID: 29704674
9)Li et al. Comparison of 22C3 PD-L1 Expression between Surgically Resected Specimens and Paired Tissue Microarrays in Non-Small Cell Lung Cancer. J Thorac Oncol. 2017;12(10):1536-1543. PMID: 28751245
10)Shen et al. Programmed cell death ligand 1 expression in osteosarcoma. Cancer Immunol Res. 2014;2(7):690-698. PMID: 24866169
11)Mimura et al. PD-L1 expression is mainly regulated by interferon gamma associated with JAK-STAT pathway in gastric cancer. Cancer Sci. 2018 ;109(1):43-53.PMID: 29034543
12)Habib et al. PDL-1 Blockade Prevents T Cell Exhaustion, Inhibits Autophagy, and Promotes Clearance of Leishmania donovani. Infect Immun. 2018;86(6). PMID: 29610255
13)Gonin et al. Expression of IL-27 by tumor cells in invasive cutaneous and metastatic melanomas. PLoS One. 2013;8(10):e75694. PMID: 24130734
14)Gagné et al. Comprehensive Assessment of PD-L1 Staining Heterogeneity in Pulmonary Adenocarcinomas Using Tissue Microarrays: Impact of the Architecture Pattern and the Number of Cores. Am J Surg Pathol. 2018;42(5):687-694.PMID: 29309297
15)Teixidó et al. Assays for predicting and monitoring responses to lung cancer immunotherapy. Cancer Biol Med. 2015;12(2):87-95. PMID: 26175924
16)Ohue et al. Survival of Lung Adenocarcinoma Patients Predicted from Expression of PD-L1, Galectin-9, and XAGE1 (GAGED2a) on Tumor Cells and Tumor-Infiltrating T Cells. Cancer Immunol Res. 2016;4(12):1049-1060. PMID: 27799141
17)Kao et al. Assessing the Effects of Concurrent versus Sequential Cisplatin/Radiotherapy on Immune Status in Lung Tumor-Bearing C57BL/6 Mice. Cancer Immunol Res. 2015;3(7):741-50. PMID: 25672395
18)Fujimura et al. Potential use of bisphosphonates in invasive extramammary Paget's disease: an immunohistochemical Investigation. Clin Dev Immunol. 2013 ;2013:164982. PMID: 23606867
19)Sakata et al. Comparison of Programmed Death Ligand-1 Immunohistochemical Staining Between Endobronchial Ultrasound Transbronchial Needle Aspiration and Resected Lung Cancer Specimens. Chest. 2018;154(4):827-837. PMID: 30059678
20)Kitazono et al. Reliability of Small Biopsy Samples Compared With Resected Specimens for the Determination of Programmed Death-Ligand 1 Expression in Non--Small-Cell Lung Cancer. Clin Lung Cancer. 2015 ;16(5):385-90.PMID: 25937270

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