Revision 2

#7963Store at +4C

1 Kit

(Reagents for 4 x 96 well plates)

Species Cross Reactivity

H Dg

UniProt ID:

#P12931

Entrez-Gene Id:

#6714

Cell Signaling Technology

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3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Volume Cap Color Storage Temp
Phospho-Src (Y416) Capture Rabbit mAb (100X) 28293 400 µl Pink +4C
Src Detection Mouse mAb (100X) 41829 400 µl Blue +4C
Anti-mouse IgG, HRP-linked Antibody (1000X) 16736 40 µl Yellow -20C

Please visit cellsignal.com for a complete listing of recommended companion products.

Description

Cell Signaling Technology's PathScan® Phospho-Src (Tyr416) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit #7953. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The phospho-Src (Tyr416) rabbit capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added followed by a total Src mouse detection antibody and anti-mouse IgG, HRP-linked antibody. HRP substrate (TMB) is then added for color development. 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.

Reagents not supplied

Phosphate Buffered Saline (PBS-20X) #9808
Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809
Cell Lysis Buffer (10X) #9803
TMB Substrate #7004
STOP Solution #7002
Blocking Buffer: 1X PBS/0.5% Tween-20, 1% BSA
96 Well Microplates**
Microplate Reader
** Antibody Pairs have been validated on Corning© 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592).

Notes: Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.

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).

  1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
  2. Hunter, T. (1987) Cell 49, 1-4.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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