Revision 3

#13598Store at -20C

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, W-S, IP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

28

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UN19

Entrez-Gene Id:

27071

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 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

DAPP1/BAM32 (D9K4O) Rabbit mAb recognizes endogenous levels of total DAPP1/BAM32 protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro159 of human DAPP1/BAM32 protein.

Background

The dual adaptor of phosphotyrosine and 3-phosphoinositides (DAPP1/BAM32) is a cytoplasmic adaptor protein that mediates the recruitment and interaction of molecules required for signal transduction downstream of the B cell receptor (BCR) (1). The DAPP1/BAM32 protein contains an amino-terminal SH2 domain and a carboxy-terminal pleckstrin homology (PH) domain that binds to PI3K-derived phosphoinositides (i.e., PIP3). Upon BCR activation, DAPP1/BAM32 is phosphorylated at specific tyrosine residues and translocated from the cytoplasm to the membrane. Research studies indicate that phosphorylation and translocation of DAPP1/BAM32 is strongly dependent upon PI3K signaling (2,3). The amino-terminal SH2 domain binds to PLCγ2 and other tyrosine-phosphorylated targets. As a result of these interactions, DAPP1/BAM32 can adjust the response to receptor activation by coordinating membrane-localized interactions among proteins of distinct signal transduction pathways (1,4). DAPP1/BAM32 is expressed most abundantly in B lymphocytes; high expression during dendritic cell (DC) maturation and localization to contact sites between DC and allogenic T cells suggest that the DAPP1/BAM32 adaptor may play a role in the activation of T cells through MHC class I-mediated signaling pathways (5).

  1. Marshall, A.J. et al. (2007) Biochem Soc Trans 35, 181-2.
  2. Marshall, A.J. et al. (2000) J Exp Med 191, 1319-32.
  3. Anderson, K.E. et al. (2000) Curr Biol 10, 1403-12.
  4. Richards, S. et al. (2008) Immunol Rev 224, 183-200.
  5. Ortner, D. et al. (2011) J Immunol 187, 3972-8.

Species Reactivity

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

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation

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 3
#13598

DAPP1/BAM32 (D9K4O) Rabbit mAb

Western Blotting Image 1: DAPP1/BAM32 (D9K4O) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using DAPP1/BAM32 (D9K4O) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: DAPP1/BAM32 (D9K4O) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (1 mg/ml) from Ramos cells DAPP1/BAM32 (D9K4O) Rabbit mAb #13598. The virtual lane view (left) shows the 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 2-40kDa.
Immunoprecipitation Image 1: DAPP1/BAM32 (D9K4O) Rabbit mAb Expand Image
Immunoprecipitation of DAPP1/BAM32 protein from Ramos cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or DAPP1/BAM32 (D9K4O) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DAPP1/BAM32 (D9K4O) Rabbit mAb.