The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 55, 60 kDa (predicted molecular weight: 55 kDa).
Use at 2-5 µg/mg of lysate.
May act in negative regulation of cell growth and transformation by interacting with nonreceptor tyrosine kinases ABL1 and/or ABL2. May play a role in regulation of EGF-induced Erk pathway activation. Involved in cytoskeletal reorganization and EGFR signaling. Together with EPS8 participates in transduction of signals from Ras to Rac. In vitro, a trimeric complex of ABI1, EPS8 and SOS1 exhibits Rac specific guanine nucleotide exchange factor (GEF) activity and ABI1 seems to act as an adapter in the complex. Regulates ABL1/c-Abl-mediated phosphorylation of ENAH. Recruits WASF1 to lamellipodia and there seems to regulate WASF1 protein level.
Widely expressed, with highest expression in brain.
Involvement in disease
Note=A chromosomal aberration involving ABI1 is a cause of acute leukemias. Translocation t(10;11)(p11.2;q23) with MLL. ABI1 isoform 2 was found to be present in acute leukemia MLL-ABI1 fusion transcript.
Belongs to the ABI family. Contains 1 SH3 domain. Contains 1 t-SNARE coiled-coil homology domain.
The t-SNARE coiled-coil homology domain is necessary and sufficient for interaction with STX1A.
In vitro substrate for v-Abl (By similarity). Phosphorylated on tyrosine residues after serum stimulation or induction by v-Abl.
Cytoplasm. Nucleus. Cell projection > lamellipodium. Cell projection > filopodium. Cell projection > growth cone. Cell junction > synapse > synaptosome. Cytoplasm > cytoskeleton. Localized to protruding lamellipodia and filopodia tips. Also localized to neuronal growth cones and synaptosomes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling SSH3BP1 with ab72318 at 1/200 (1µg/ml). Detection: DAB.
Western blot - SSH3BP1 antibody (ab72318)
All lanes : Anti-SSH3BP1 antibody (ab72318) at 0.04 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg
Detection of Human SSH3BP1 by Western Blot of Immunoprecipitates.
Samples: Whole cell lysate (1 mg for IP, 20% of IP loaded) from HeLa cells.
Lane 1: ab72317 used for IP at 3 µg/mg lysate.
Lane 2: SSH3BP1 immunoprecipitated with an other antibody.
Lane 3: ab72318 at 3 µg/mg lysate used for IP of SSH3BP1, which recognize downstream epitopes.
Lane 4: control Ig IP
For blotting immunoprecipitated SSH3BP1, ab72318 was used at 1 µg/ml.
Immunoprecipitation - SSH3BP1 antibody (ab72318)
Detection of SSH3BP1 by Immunoprecipitation from Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab72318 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent WB detection. Lane 2 represents control IgG IP.