Cell Signaling Technology

Product Pathways - CD Markers

PathScan® Total PD-L1 Sandwich ELISA Kit #14784

B7H1   CD274   ELISA   PD-L1   sandwich elisa   screen  

REACTIVITY
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No. Size Price
14784C 1 Kit ( 96 assays ) ¥7,500.00 现货查询 购买询价
Product Includes Volume Solution Color
ELISA Sample Diluent 25 ml Blue
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
PD-L1 Mouse Detection mAb 1 ea Green (Lyophilized)
PD-L1 Rabbit mAb Coated Microwells 96 tests
STOP Solution #7002 11 ml Colorless
TMB Substrate #7004 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
Sealing Tape 2 sheets

Specificity / Sensitivity

PathScan® Total PD-L1 Sandwich ELISA Kit detects endogenous levels of PD-L1 protein as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Total PD-L1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PD-L1 protein. A PD-L1 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, PD-L1 protein is captured by the coated antibody. Following extensive washing, a PD-L1 mouse detection antibody is added to detect the captured PD-L1 protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of PD-L1 protein.

Antibodies in kit are custom formulations specific to kit.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. PD-L1 protein is detected from KARPAS-299 and SUP-M2 cell lines using the PathScan® Total PD-L1 Sandwich ELISA Kit #14784. The absorbance readings at 450 nm are shown in the top figure, while corresponding western blots using PD-L1 (E1L3N®) XP® Rabbit mAb #13684 are shown in the bottom figure. Cell Line Source: Dr. Abraham Karpas at the University of Cambridge.

Sensitivity

Sensitivity

Figure 2. The relationship between protein concentration of lysates from SUP-M2 cells and the absorbance at 450 nm as detected by the PathScan® Total PD-L1 Sandwich ELISA Kit #14784 is shown. Unstarved SUP-M2 cells (0.5-1.0 x 106) were harvested and then lysed.

Background

Programmed cell death 1 ligand 1 (PD-L1, B7-H1, CD274) is a member of the B7 family of cell surface ligands that regulate T cell activation and immune responses. The PD-L1 ligand binds the PD-1 transmembrane receptor and inhibits T cell activation. PD-L1 was discovered following a search for novel B7 protein homologs and was later shown to be expressed by antigen presenting cells, activated T cells, and tissues including placenta, heart, and lung (1-3). Similar in structure to related B7 family members, PD-L1 protein contains extracellular IgV and IgC domains and a short, cytoplasmic region. Research studies demonstrate that PD-L1 is expressed in several tumor types, including melanoma, ovary, colon, lung, breast, and renal cell carcinomas (4-6). Expression of PD-L1 in cancer is associated with tumor infiltrating lymphocytes, which mediate PD-L1 expression through the release of interferon gamma (7). Additional research links PD-L1 expression to cancers associated with viral infections (8,9).

  1. Dong, H. et al. (1999) Nat Med 5, 1365-9.
  2. Freeman, G.J. et al. (2000) J Exp Med 192, 1027-34.
  3. Liang, S.C. et al. (2003) Eur J Immunol 33, 2706-16.
  4. Dong, H. et al. (2002) Nat Med 8, 793-800.
  5. Thompson, R.H. et al. (2006) Cancer Res 66, 3381-5.
  6. Pardoll, D.M. (2012) Nat Rev Cancer 12, 252-64.
  7. Taube, J.M. et al. (2012) Sci Transl Med 4, 127ra37.
  8. Lyford-Pike, S. et al. (2013) Cancer Res 73, 1733-41.
  9. Chen, B.J. et al. (2013) Clin Cancer Res 19, 3462-73.

Application References

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Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

PathScan is a trademark of Cell Signaling Technology, Inc.

E1L3N is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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