Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

PathScan® Total B7-H4 Sandwich ELISA Kit #14520

B7-H4   B7H4   elisa   sandwich elisa   screen   VTCN-1   VTCN1  

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No. Size Price
14520C 1 Kit ( 96 assays ) ¥6,338.00 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
B7-H4 Mouse Detection mAb 1 ea Green (Lyophilized)
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
B7-H4 Rabbit mAb Coated Microwells 96 tests
Sealing Tape 2 sheets

Specificity / Sensitivity

PathScan® Total B7-H4 Sandwich ELISA Kit detects endogenous levels of B7-H4 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

PathScan® Total B7-H4 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of B7-H4. A B7-H4 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, B7-H4 protein is captured by the coated antibody. Following extensive washing, a B7-H4 Mouse Detection mAb is added to detect the captured B7-H4 protein. A HRP-linked, anti-mouse antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total B7-H4.

Antibodies in kit are custom formulations specific to kit.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. B7-H4 protein is expressed in SK-BR-3 cells but absent in H1299 cells, as detected using the PathScan® Total B7-H4 Sandwich ELISA Kit #14520. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using B7-H4 (D1M8I) XP® Rabbit mAb #14572 is shown in the bottom figure.

Sensitivity

Sensitivity

Figure 2. The relationship between protein concentration of lysates from SK-BR-3 cells and the absorbance at 450 nm as detected by the PathScan® Total B7-H4 Sandwich ELISA Kit #14520 is shown. SK-BR-3 cells (85% confluence) were harvested and then lysed.

Background

B7 homolog 4 (B7-H4, VTCN1) is a member of the B7 family of cell surface ligands that regulate T cell activation and immune responses (1-3). B7-H4 protein contains two extracellular Ig-like V-type domains, a transmembrane domain, and a short, two amino acid intracellular domain (3). The B7-H4 protein is shown to inhibit T cell activation, proliferation, and cytokine production (1,4,5). Although B7-H4 mRNA is widely expressed, B7-H4 protein is restricted to antigen presenting cells and B cells (1). The B7-H4 protein is also found in several tumor types, including ovarian cancer and breast cancer (6). Research studies indicate that B7-H4 protein is present on the surface of ovarian tumor cells, and that targeted inhibition of B7-H4 using recombinant antibodies restores T cell activation pathways. These studies suggest some potential therapeutic value in blocking B7-H4 function and restoring T cell function in cancer patients (7,8).

  1. Sica, G.L. et al. (2003) Immunity 18, 849-61.
  2. Prasad, D.V. et al. (2003) Immunity 18, 863-73.
  3. Zang, X. et al. (2003) Proc Natl Acad Sci U S A 100, 10388-92.
  4. He, C. et al. (2011) Clin Dev Immunol 2011, 695834.
  5. Yi, K.H. and Chen, L. (2009) Immunol Rev 229, 145-51.
  6. Salceda, S. et al. (2005) Exp Cell Res 306, 128-41.
  7. Dangaj, D. and Scholler, N. (2013) Oncoimmunology 2, e25913.
  8. Dangaj, D. et al. (2013) Cancer Res 73, 4820-9.

Application References

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Protocols

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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.

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

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