Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P14317

Entrez-Gene Id:

3059

Product Information

Product Usage Information

Application Dilution
Immunofluorescence (Immunocytochemistry) 1:50
Flow Cytometry (Fixed/Permeabilized) 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb recognizes endogenous levels of HS1 protein only when phosphorylated at Tyr378 or Tyr397.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr405 of mouse HS1 protein. This site corresponds to Tyr397 of human HS1 protein.

Background

HS1 (HCLS1, LckBP1, p75) is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (1,2). HS1 contains four cortactin repeats and a single SH3 domain (2). This intracellular protein is phosphorylated following immune receptor activation, which promotes recruitment of HS1 to the immune synapse (3-5). Phosphorylation of HS1 is required to regulate actin dynamics and provide docking sites for many other signaling molecules, such as Vav1 and PLCγ1 (6). HS1 also plays an important role in platelet activation (7).
HS1 is rapidly phosphorylated at Tyr397 by Syk and/or Lyn kinases following immune receptor stimulation and thrombin-mediated platelet stimulation. This phosphorylation is an important step in cytoskeletal rearrangement and signaling complex formation (6-10).

  1. Kitamura, D. et al. (1989) Nucleic Acids Res 17, 9367-79.
  2. Kitamura, D. et al. (1995) Biochem Biophys Res Commun 208, 1137-46.
  3. Suzuki, H. et al. (1997) J Immunol 159, 5881-8.
  4. Hata, D. et al. (1994) Immunol Lett 40, 65-71.
  5. Yamanashi, Y. et al. (1993) Proc Natl Acad Sci USA 90, 3631-5.
  6. Gomez, T.S. et al. (2006) Immunity 24, 741-52.
  7. Kahner, B.N. et al. (2007) Blood 110, 2449-56.
  8. Yamanashi, Y. et al. (1997) J Exp Med 185, 1387-92.
  9. Hao, J.J. et al. (2004) J Biol Chem 279, 33413-20.
  10. Brunati, A.M. et al. (2005) J Biol Chem 280, 21029-35.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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.
XP 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.

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 6
#8714

Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb

Immunofluorescence Image 1: Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of Ramos cells, untreated (left) or treated with IgM (12 μg/mL, 10 min, right) using Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb Expand Image
Flow cytometric analysis of Ramos cells, untreated (blue) or IgM-treated (green), using Phospho-HS1 (Tyr378/397) (D12C1) XP® Rabbit mAb #8714. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 conjugate) #4412 was used as a secondary antibody.