Product Pathways - Tyrosine Kinase/ Adaptors
Src Antibody Sampler Kit #9935
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Isotype |
|---|---|---|---|---|---|
| Non-phospho-Src (Tyr527) Antibody # 2107 | 40 microliters | W | H M R (C) | 60 | Rabbit |
| Non-phospho-Src (Tyr416) (7G9) Mouse mAb # 2102 | 40 microliters | W IP | H M R (C) (X) | 60 | Mouse IgG2b |
| Phospho-Src (Tyr527) Antibody # 2105 | 40 microliters | W IHC-P | H M R (C) | 60 | Rabbit |
| Phospho-Src Family (Tyr416) Antibody # 2101 | 40 microliters | W | H M R (C) (X) | 60 | Rabbit |
| Src (36D10) Rabbit mAb # 2109 | 40 microliters | W IP IHC-P IHC-F IF-F IF-IC F | H M R Mk Pg Hm B (C) | 60 | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 100 microliters | Goat | |||
| Anti-mouse IgG, HRP-linked Antibody # 7076 | 100 microliters | Horse |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
IF-F=Immunofluorescence (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Pg=Pig
C=Chicken
Hm=Hamster
B=Bovine
X=Xenopus
Specificity / Sensitivity
Each antibody in the kit detects endogenous levels of Src only when in the indicated phosphorylation state at the indicated residue. The antibodies may cross-react with other Src family members when in equivalent phosphorylation states.
Western Blotting
Western blot analysis of extracts from serum-starved COLO 201 cells, v-Src antibody immunoprecipitates from COLO 201 cells and PDGF-treated NIH/3T3 cells, treated with λ phosphatase as indicated, using Phospho-Src (Tyr416) Antibody #2101 (upper) or v-Src antibody (lower).
Western Blotting
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 ) mAb #2102 (upper), v-Src antibody (middle) or Phospho-Src (Tyr416) Antibody #2101 (lower).
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells, using Phospho-Src (Tyr527) Antibody #2105 (A, B) or v-Src antibody (C, D). The phospho-specificity of the antibody was determined by treating the membrane with (B, D) or without (A, C) calf intestinal alkaline phosphatase (CIP) after Western transfer.
Western Blotting
Western blot analysis of extracts from A431 cells, untreated or pervanidate-treated (1 mM, 15 minutes), using Non-phospho-Src (Tyr527) Antibody #2107 (upper) or v-Src antibody (lower).
Western Blotting
Western blot analysis of extracts from various cell lines using Src (36D10) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing membrane localization of phosphorylated Src, using Phospho-Src (Tyr527) Antibody #2105.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Src (36D10) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
IF-IC
Immunofluorescent staining of MCF-7 cells, using Src (36D10) Rabbit mAb #2109 alone (A), or the same antibody after a 1 hour preincubation with antigenic peptide (B).
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 four Western blot experiments per primary antibody.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic phospho- or non-phosphopeptides corresponding to residues surrounding Tyr416 or Tyr527 of human Src protein. The mouse monoclonal antibody is produced by immunizing animals with a synthetic non-phosphopeptide corresponding to residues surrounding Tyr416 of human Src protein. The rabbit monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy-terminus of human Src. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
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. Phosphorylation of Src at Tyr416 in the activation loop of the kinase domain by carboxy-terminal Src kinase (Csk) upregulates enzyme activity, whereas phosphorylation of Tyr527 in the carboxy-terminal tail 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).
- 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. USA 86, 6538-6542.
- Yamanashi, Y. et al. (1991) Science 251, 192-194.
- Molina, T. J. et al. (1992) Nature 357, 161-164.
- Straus, D. B. et al. (1992) Cell 70, 585-593.
- Chow, L. M. et al. (1993) Nature 365, 156-160.
Application References
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Protocols
- 2101:
- Western Blotting
- 2102:
- Immunoprecipitation, Western Blotting
- 2105:
- IHC / Paraffin, Western Blotting
- 2107:
- Western Blotting
- 2109:
- Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
Companion Products
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
Rabbit Monoclonals Produced UsingTechnology from Epitomics, Inc. UnderU.S. patent No. 5,675,063.
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.
