|Application:||IHC-P, IHC-F, WB|
Data & Images
|Product Name||Anti-Alpha-Actinin Antibody (Monoclonal, EA-53)|
|Description||Mouse IgG monoclonal antibody for alpha-Actinin, actinin, alpha 2 (ACTN2 ) detection. Tested with WB, IHC-P, IHC-F in Human;mouse;rat;rabbit. No cross reactivity with other proteins.|
|Cite This Product||Anti-Alpha-Actinin Antibody (Monoclonal, EA-53) EA-53 (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1104)|
|Replacement Item||This antibody may replace the following items: sc-130928|sc-15335|sc-166524|sc-17829|sc-7453|sc-7454-R from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rabbit, Rat|
|Predicted Species||Monkey, Rabbit|
*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.
|Application||IHC-P, IHC-F, WB
*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-Mouse IgG (EK1001) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (SV0001-1) for IHC(P) and IHC(F).
*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 2017!
|Immunogen||Rabbit skeletal alpha-actinin.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.
*carrier free antibody available upon request.
|Concentration||Add 1ml of PBS buffer will yield a concentration of 100ug/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.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Molecular Weight||103834 MW|
|Protein Function||ACTN2: F-actin cross-linking protein which is thought to anchoractin to a variety of intracellular structures. This is a bundlingprotein.|
|Tissue Specificity||ACTN2: Expressed in both skeletal and cardiac muscle.|
|Sequence Similarities||Belongs to the alpha-actinin family.|
|Subcellular Localization||ACTN2: Cytoplasm, myofibril, sarcomere, Z line
Actn2: Cytoplasm, myofibril, sarcomere, Z line
|Alternative Names||Protein Actn2 ;RCG30552 ;Actn2 ;rCG_30552 ;|
|Research Areas|||signal transduction|cytoskeleton / ecm|cytoskeleton|microfilaments|actin etc|actin binding proteins| cardiovascular|heart|cardiac arrhythmias|contractility|contractile proteins|actins||
Background for Alpha-actinin-2
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Alpha-Actinin Antibody (Monoclonal, EA-53) 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), 1-2μg/ml, Human, mouse, rat, rabbit, By Heat
Immunohistochemistry(Frozen Section), 1-2μg/ml, Human, mouse, rat, rabbit, -
Western blot, 0.5-1μg/ml, Human, mouse, rat, rabbit
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Alpha-Actinin Antibody (Monoclonal, EA-53) 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: rat cardiac muscle tissue lysates,
Lane 2: rat skeletal muscle tissue lysates,
Lane 3: mouse cardiac muscle tissue lysates,
Lane 4: mouse skeletal muscle tissue lysates,
Lane 5: human placenta tissue lysate.
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 mouse anti- Alpha-Actinin antigen affinity purified monoclonal antibody (Catalog # MA1104) 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 Biotin Conjugated goat anti-mouse IgG 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 # EK1001) with Tanon 5200 system. A specific band was detected for Alpha-Actinin at approximately 103KD. The expected band size for Alpha-Actinin is at 103KD.
Alpha-Actinin was detected in paraffin-embedded section of mouse lung 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 1.5μg/ml mouse anti- Alpha-Actinin Antibody (MA1104) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Alpha-Actinin was detected in paraffin-embedded section of human mammary 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 1.5μg/ml mouse anti- Alpha-Actinin Antibody (MA1104) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.