Product Pathways - PathScan ELISA
PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit #7953
|7953S||1 Kit (96 assays)||---||In Stock||---|
|7953||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.
|Kit Includes||Volume||Solution Color|
|Phospho-Src (Tyr416) Rabbit Antibody Coated Microwells||96 tests|
|Src Mouse Detection Antibody||11 ml||Green|
|Anti-mouse 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 Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Src (Tyr416). A phospho-Src rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-Src (Tyr416) is captured by the coated antibody. Following extensive washing, a Src mouse detection antibody is added to detect the captured phospho-Src (Tyr416). Anti-mouse, HRP-linked antibody is then used to recognize the bound detection antibody. The HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Src (Tyr416).
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
The PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of Src when phosphorylated at Tyr416, as shown in Figure 1. This kit does not cross-react with the Src family members Yes, Lck, Lyn or Hck. Other family members have not been tested. 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.
Figure 2. The relationship between lysate protein concentration from untreated and H202-treated MDCK cells and the absorbance at 450 nm is shown. MDCK cells were treated with H202 (3 mM) for 3 minutes at 37ºC and then lysed.
ELISA - Western correlation
Figure 1. Phosphorylation of Src at Tyr416 in H202 stimulated MDCK cells (3 mM for 3 min) and untreated Calu-3 cells is detected by the PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit. In contrast, a low level of phospho-Src (Tyr416) is detected in untreated MDCK cells and Calu-3 cells lysed in the presence of λ-phosphatase. The absorbance readings at 450 nm are shown in the top figure while western blots, using Phospho-Src Family (Tyr416) (100F9) Rabbit mAb #2113 (upper) or Src (L4A1) Mouse mAb #2110 (lower), are shown in the bottom figure.
The Src family of protein tyrosine kinases, which includes Src, Lyn, Fyn, Yes, Lck, Blk, and Hck, are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk renders the enzyme less active (2).
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- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7984 PathScan® Total Src Sandwich ELISA Kit
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 2101 Phospho-Src Family (Tyr416) Antibody
- 7002 STOP Solution
- 2109 Src (36D10) Rabbit mAb
- 7004 TMB Substrate
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.