|Validated Species:||Human, Mouse, Rat|
Data & Images
|Product Name||Anti-Alpha Amylase 1 Antibody|
|Description||Rabbit IgG polyclonal antibody for Alpha-amylase 1(AMY1A) detection. Tested with WB, IHC-P in Human;Mouse;Rat.|
|Cite This Product||Anti-Alpha Amylase 1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1486)|
|Replacement Item||This antibody may replace the following items: sc-12821|sc-166349|sc-25562|sc-31868|sc-31869|sc-31870|sc-46657|sc-514229|sc-514313 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||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).
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human Alpha Amylase 1(30-44aa HLFEWRWVDIALECE), different from the related rat and mouse sequences by one amino acid.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||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.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Alpha-amylase 1|
|Molecular Weight||57768 MW|
|Sequence Similarities||Belongs to the glycosyl hydrolase 13 family.|
|Alternative Names||Alpha-amylase 1;220.127.116.11;1,4-alpha-D-glucan glucanohydrolase 1;Salivary alpha-amylase;AMY1A;AMY1;AMY1B;AMY1;AMY1C;AMY1;|
Background for Alpha-amylase 1
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Alpha Amylase 1 Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Rat, Human, Mouse, By Heat|
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Alpha Amylase 1 Antibody Images
Click the images to enlarge.
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: human PANC-1 whole cell lysates,
Lane 2: rat pancreas tissue lysates,
Lane 3: mouse pancreas 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- Alpha Amylase 1 antigen affinity purified polyclonal antibody (Catalog # PA1486) at 0.5 μg/mL overnight at 4°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 Alpha Amylase 1 at approximately 55-58KD. The expected band size for Alpha Amylase 1 is at 58KD.
Alpha Amylase 1 was detected in paraffin-embedded section of Rat Spleen 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 1μg/ml rabbit anti- Alpha Amylase 1 Antibody (PA1486) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
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.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,