|Reactivity||Human, Mouse, Rat|
|Applications||IF, IHC-P, WB|
|Product Name||Anti-SSH3BP1/ABI1 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Abl interactor 1(ABI1) detection. Tested with WB, IHC-P, IF in Human;Mouse;Rat.|
|Cite This Product||Anti-SSH3BP1/ABI1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1001)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human ABI1 (483-494aa WYEGVCNRVTGL), different from the related rat and mouse sequences by one amino acid.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Immunofluorescence, 2μg/ml, Human, -
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 2. IF analysis of ABI using anti- ABI antibody (PA1001)
ABI was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1Î¼g/mL rabbit anti- ABI Antibody (PA1001) overnight at 4Â°C. DyLightÂ®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37Â°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 3. IHC analysis of ABI using anti-ABI antibody (PA1001).
ABI was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1Î¼g/ml rabbit anti-ABI Antibody (PA1001) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 4. IHC analysis of ABI using anti-ABI antibody (PA1001).
ABI was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1Î¼g/ml rabbit anti-ABI Antibody (PA1001) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 1. Western blot analysis of ABI-1 using anti- ABI-1 antibody (PA1001).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates,
Lane 3: human THP-1 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- ABI-1 antigen affinity purified polyclonal antibody (Catalog # PA1001) at 0.5 Î¼g/mL overnight at 4Â°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ABI-1 at approximately 65KD. The expected band size for ABI-1 is at 55KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Abl interactor 1|
|Tissue Specificity||Widely expressed, with highest expression in brain. .|
|Alternative Names||Abl interactor 1;Abelson interactor 1;Abi-1;Abl-binding protein 4;AblBP4;Eps8 SH3 domain-binding protein;Eps8-binding protein;Nap1-binding protein;Nap1BP;Spectrin SH3 domain-binding protein 1;e3B1;ABI1;SSH3BP1;|
|Subcellular Localization||Cytoplasm . Nucleus . Cell projection, lamellipodium . Cell projection, filopodium . Cell projection, growth cone . Cell junction, synapse, postsynaptic cell membrane, postsynaptic density . Cytoplasm, cytoskeleton . Localized to protruding lamellipodia and filopodia tips. Also localized to neuronal growth cones and synaptosomes. May shuttle from the postsynaptic densities to the nucleus (By similarity). .|
|Molecular Weight||55081 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||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. In brain, seems to regulate the dendritic outgrowth and branching as well as to determine the shape and number of synaptic contacts of developing neurons. .|
|Research Areas||Human, Mouse, Rat
*You can search these to find other products in these research areas.
|Background||ABI1 is a human homolog of mouse Abl-interactor-1(Abi1), mapped on 10p11.2. ABI1 participates in the transduction of signals from Ras to Rac by regulating Rac-specific guanine nucleotide exchange factor(GEF) activities. ABI1 dramatically promoted ABL1-mediated tyrosine phosphorylation of MENA, but not of other substrates. Abi-1 regulates c-Abl-mediated phosphorylation of Mena by interacting with both proteins. ABI1 plays a role in the leukemogenesis by translocating to MLL.|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at firstname.lastname@example.org for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact email@example.com
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?A: Some common names include but are not limited to nap1 antibody, abi1 antibody