Product Pathways - Lymphocyte Signaling
PathScan® Total B7-H4 Sandwich ELISA Kit #14520
|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||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.
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
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.
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).
- Sica, G.L. et al. (2003) Immunity 18, 849-61.
- Prasad, D.V. et al. (2003) Immunity 18, 863-73.
- Zang, X. et al. (2003) Proc Natl Acad Sci U S A 100, 10388-92.
- He, C. et al. (2011) Clin Dev Immunol 2011, 695834.
- Yi, K.H. and Chen, L. (2009) Immunol Rev 229, 145-51.
- Salceda, S. et al. (2005) Exp Cell Res 306, 128-41.
- Dangaj, D. and Scholler, N. (2013) Oncoimmunology 2, e25913.
- Dangaj, D. et al. (2013) Cancer Res 73, 4820-9.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 14572 B7-H4 (D1M8I) XP® Rabbit mAb
- 7004 TMB Substrate
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9998 BSA
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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