Product Pathways - PathScan ELISA
PathScan® Total Zap-70 Sandwich ELISA Kit #7172
|7172S||1 Kit (96 assays)||---||In Stock||---|
|7172||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
Important Ordering Details: Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
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|Kit Includes||Volume||Solution Color|
|Zap-70 mAb Coated Microwells||96 tests|
|Zap-70 Detection Ab||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
CST's PathScan® Total Zap-70 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Zap-70 protein. A Zap-70 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-Zap-70 proteins are captured by the coated antibody. Following extensive washing, Zap-70 Antibody is added to detect the captured phospho- and nonphospho-Zap-70 protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total Zap-70 protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Total Zap-70 Sandwich ELISA Kit detects endogenous levels of Zap-70 enzyme. As shown in Figure 1, using this Sandwich ELISA Kit #7171, a significant induction of Phospho-Zap-70 (Tyr319) in Jurkat cells treated with hydrogen peroxide is detected. However, the level of Zap-70 (either untreated or treated), detected by the Total Zap-70 Sandwich ELISA Kit #7172, remains unchanged. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of Jurkat cells with hydrogen peroxide stimulates phopsphorylation of Zap-70 at Tyr319, detected by PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA kit #7171, but does not affect the level of total Zap-70 detected by PathScan® Total Zap-70 Sandwich ELISA kit #7172. OD 450 readings are shown in the top figure, while the corresponding Western blot using Phospho-Zap-70 (Tyr319) Ab #2701 (right panel) or Zap-70 Rabbit mAb #2705 (left panel), is shown in the bottom figure.
The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).
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For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.