Revision 1
#9935
Store at -20C
Src Antibody Sampler Kit
1 Kit
(5 x 20 microliters)
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
---|---|---|---|---|
Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb | 6943 | 20 µl | 60 kDa | Rabbit IgG |
Non-phospho-Src (Tyr527) Antibody | 2107 | 20 µl | 60 kDa | Rabbit |
Non-phospho-Src (Tyr416) (7G9) Mouse mAb | 2102 | 20 µl | 60 kDa | Mouse IgG2b |
Phospho-Src (Tyr527) Antibody | 2105 | 20 µl | 60 kDa | Rabbit |
Src (32G6) Rabbit mAb | 2123 | 20 µl | 60 kDa | Rabbit IgG |
Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat | |
Anti-mouse IgG, HRP-linked Antibody | 7076 | 100 µl | Horse |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
Description
The Src Antibody Sampler kit provides an economical means of evaluating total Src protein levels and its phosphorylation status. The kit contains enough primary and secondary antibodies to perform two Western blot experiments per primary antibody.
Storage
Background
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).
Lyn is a member of the Src family that is predominantly expressed in hematopoietic cells (3). Lyn participates in signaling from multiple cell surface receptors such as the B cell Ag receptor (BCR) and CD40 (4).
Lck is essential for T-lymphocyte activation and differentiation (5,6). The activity of Lck is regulated by protein kinases and phosphatases. Phosphorylation of the C-terminal tyrosine 505 serves to downregulate Lck catalytic activity, while phosphorylation at tyrosine 394 leads to an increase in Lck activity (7).
Background References
- Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
- Hunter, T. (1987) Cell 49, 1-4.
- Yamanashi, Y. et al. (1989) Proc Natl Acad Sci U S A 86, 6538-42.
- Yamanashi, Y. et al. (1991) Science 251, 192-4.
- Molina, T.J. et al. (1992) Nature 357, 161-4.
- Straus, D.B. and Weiss, A. (1992) Cell 70, 585-93.
- Chow, L.M. et al. (1993) Nature 365, 156-60.
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 1
#9935
Src Antibody Sampler Kit
Western blot analysis of extracts from pervanadate-treated (1 mM for 5 minutes) NIH/3T3 cells that have been stably transfected with a constitutively active form of Src (in which the regulatory tyrosine 527 residue has been mutated to phenylalanine), A431 cells and HepG2 cells, using Non-phospho-Src (Tyr416) (7G9 ) Mouse mAb (top), v-Src antibody (middle) or Phospho-Src (Tyr416) Antibody #2101 (bottom).
Simple Western™ analysis of lysates (1 mg/mL) from HepG2 cells using Non-phospho-Src (Tyr416) (7G9) Mouse mAb #2102. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from NIH/3T3 cells, using Phospho-Src (Tyr527) Antibody (A,B) or v-Src antibody (C,D). The phospho-specificity of the antibody was confirmed by treating the membrane with calf intestinal alkaline phosphatase (CIP) (B,D) after Western transfer.
Revision 1
#9935
Src Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from 3T3 cells using Phospho-Src (Tyr527) Antibody #2105. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from A431 cells, untreated or pervanadate-treated (1 mM for 15 minutes), using Non-phospho-Src (Tyr527) Antibody (upper) or v-Src antibody (lower).
Western blot analysis of cell extracts from various cell lines, using Src (32G6) Rabbit mAb.
Revision 1
#9935
Src Antibody Sampler Kit
Western blot analysis of extracts NIH/3T3 cells, serum-starved or treated with human Platelet-Derived Growth Factor BB hPDGF-BB #8912 (100 ng/ml, 15 min), using Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb (upper) or Src (36D10) Rabbit mAb #2109 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Revision 1
#9935
Src Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing membrane and cytoplasmic localization using Phospho-Src (Tyr527) Antibody.