|Product Name||Anti-IGFBP3 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 Insulin-like growth factor-binding protein 3(IGFBP3) detection. Tested with WB in Human;Rat.|
|Cite This Product||Anti-IGFBP3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1498)|
|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 in the middle region of human IGFBP-3(182-198aa IKKGHAKDSQRYKVDYE).|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human, Rat
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.
Images And Assay Conditions
Anti-IGFBP-3 antibody, PA1498, Western blotting
All lanes: Anti IGFBP-3 (PA1498) at 0.5ug/ml
Lane 1: 293T Whole Cell Lysate at 40ug
Lane 2: MCF- 7 Whole Cell Lysate at 40ug
Lane 3: A549 Whole Cell Lysate at 40ug
Lane 4: SW620 Whole Cell Lysate at 40ug
Predicted bind size: 32KD
Observed bind size: 40KD
Figure 2. Western blot analysis of IGFBP3 using anti- IGFBP3 antibody (PA1498).
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 liver tissue lysates,
Lane 2: rat kidney tissue lysates,
Lane 3: rat heart tissue 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- IGFBP3 antigen affinity purified polyclonal antibody (Catalog # PA1498) 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 IGFBP3 at approximately 40KD. The expected band size for IGFBP3 is at 32KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Insulin-like growth factor-binding protein 3|
|Tissue Specificity||Expressed by most tissues. Present in plasma.|
|Alternative Names||Insulin-like growth factor-binding protein 3;IBP-3;IGF-binding protein 3;IGFBP-3;IGFBP3;IBP3;|
|Subcellular Localization||Secreted .|
|Molecular Weight||31674 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors. Also exhibits IGF-independent antiproliferative and apoptotic effects mediated by its receptor TMEM219/IGFBP-3R. .|
|Research Areas||Human, Rat
*You can search these to find other products in these research areas.
|Background||IGFBP3, Insulin-like growth fator-binding protein 3, is a member of the insulin-like growth factor binding protein(IGFBP) family and encodes a protein with an IGFBP domain and a thyroglobulin type-I domain. IGFBP3 is located on chromosome 7. The protein forms a ternary complex with insulin-like growth factor acid-labile subunit(IGFALS) and either insulin-like growth factor(IGF) I or II. In this form, it circulates in the plasma, prolonging the half-life of IGFs and altering their interaction with cell surface receptors. Alternate transcriptional splice variants, encoding different isoforms, have been characterized.|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at email@example.com 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 firstname.lastname@example.org
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 igfbp 3 antibody, igfbp3 antibody, igfbp-3 antibody