Revision 7
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:

WB, IHC-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

85 (human, monkey), 100 (rat), 120 (murine isoform 1), 55 (murine isoform 2)

Source/Isotype:

Rabbit IgG

UniProt ID:

#P78324

Entrez-Gene Id:

140885

Product Information

Product Usage Information

This product is the carrier free version of product #13379. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN, or CUT&Tag assays. If you require a carrier-free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #13379.

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total SHPS1 protein. This antibody recognizes both large and small isoforms of murine mSHPS1/SIRPα.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro413 of human SIRPα/SHPS1 protein.

Background

SHP-substrate 1 (SHPS1, SIRPα) is a single-pass membrane protein and member of both the immunoglobulin superfamily and the signal regulatory protein (SIRP) family. Following growth hormone stimulation or integrin binding, SHPS1 is phosphorylated at several tyrosine residues within its cytoplasmic tail. These phosphorylation events promote association between SHPS1 and multiple signaling proteins, including SHP-1, SHP-2, Grb2 and Shc via their SH2 domains (1-4). Recruitment of SHP-1 and SHP-2 results in SHPS1 dephosphorylation and suppression of tyrosine kinase signaling (1-3,5). The tyrosine kinase JAK2 associates with SHPS1 via its carboxy terminus and phosphorylates SHPS1 in response to extracellular stimuli (5). Research studies show that Src associates with and may phosphorylate SHPS1 in response to insulin (4). In macrophages, SHPS1 can form a complex with the Src pathway adaptor protein SKAP2, Fyn-binding protein FYB, and the tyrosine kinase PYK2 (6). The SHPS1 extracellular domain contains at least three IgG-like domains that interact with CD47, a ubiquitously expressed, integrin-associated protein that acts as a repressive cue in both immune and neuronal cells (7,8). The interaction between CD47 and SHPS1 on opposing cells can inhibit cellular migration (9), promote "tethering" between macrophages and target cells during engulfment (10), facilitate self versus non-self recognition (11), and maintain immune homeostasis (12). SHPS1 plays a critical role in modulating the immune response and inflammation, and may play a role in neuronal development (13,14). The interaction between SHPS1 and CD47 may be an exploitable target in cancer therapy (15-17).

  1. Kharitonenkov, A. et al. (1997) Nature 386, 181-6.
  2. Ochi, F. et al. (1997) Biochem Biophys Res Commun 239, 483-7.
  3. Takada, T. et al. (1998) J Biol Chem 273, 9234-42.
  4. Shen, X. et al. (2009) Mol Cell Proteomics 8, 1539-51.
  5. Stofega, M.R. et al. (2000) J Biol Chem 275, 28222-9.
  6. Timms, J.F. et al. (1999) Curr Biol 9, 927-30.
  7. Seiffert, M. et al. (1999) Blood 94, 3633-43.
  8. Vernon-Wilson, E.F. et al. (2000) Eur J Immunol 30, 2130-7.
  9. Motegi, S. et al. (2003) EMBO J 22, 2634-44.
  10. Tada, K. et al. (2003) J Immunol 171, 5718-26.
  11. van Beek, E.M. et al. (2005) J Immunol 175, 7781-7.
  12. Legrand, N. et al. (2011) Proc Natl Acad Sci U S A 108, 13224-9.
  13. Sarfati, M. et al. (2008) Curr Drug Targets 9, 842-50.
  14. Matozaki, T. et al. (2009) Trends Cell Biol 19, 72-80.
  15. Hara, K. et al. (2011) Cancer Res 71, 1229-34.
  16. Willingham, S.B. et al. (2012) Proc Natl Acad Sci U S A 109, 6662-7.
  17. Weiskopf, K. et al. (2013) Science 341, 88-91.

Species Reactivity

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

Applications Key

WB: Western Blotting IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) 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.

使用に関する制限

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Revision 7
#47027

SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Western blot analysis of extracts from various cell lines using SIRPα/SHPS1 (D6I3M) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded breast ductal carcinoma using SIRPa/SHPS1 (D6I3M) Rabbit mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using SIRPa/SHPS1 (D6I3M) Rabbit mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded mouse lung using SIRPa/SHPS1 (D6I3M) Rabbit mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using SIRPα/SHPS1 (D6I3M) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human lymph node using SIRPα/SHPS1 (D6I3M) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded ACHN (left) or MCF7 (right) cell pellets using SIRPα/SHPS1 (D6I3M) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Confocal immunofluorescent analysis of ACHN (left, high-expressing) and MCF7 (right, low-expressing) cells using SIRPα/SHPS1 (D6I3M) Rabbit mAb (green) and DRAQ5® #4084 (fluorescent DNA dye) (blue). Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: SIRPα/SHPS1 (D6I3M) Rabbit mAb (BSA and Azide Free) Expand Image
Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and U937 cells (green, positive) using SIRPα/SHPS1 (D6I3M) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data were generated using the standard formulation of this antibody.