Product Info Summary
SKU: | A00994-3 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-AMPK alpha 1/PRKAA1 Antibody Picoband™
View all AMPK alpha 1 Antibodies
SKU/Catalog Number
A00994-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AMPK alpha 1/PRKAA1 Antibody Picoband™ catalog # A00994-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-AMPK alpha 1/PRKAA1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00994-3)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human AMPK alpha 1 recombinant protein (Position: F377-R446).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00994-3 is reactive to PRKAA1 in Human, Mouse, Rat
Applications
A00994-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Observed Molecular Weight
63 kDa
Calculated molecular weight
Background of AMPK alpha 1
5'-AMP-activated protein kinase catalytic subunit alpha-1 is an enzyme that in humans is encoded by the PRKAA1 gene. The protein encoded by this gene belongs to the ser/thr protein kinase family. It is the catalytic subunit of the 5'-prime-AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensor conserved in all eukaryotic cells. The kinase activity of AMPK is activated by the stimuli that increase the cellular AMP/ATP ratio. AMPK regulates the activities of a number of key metabolic enzymes through phosphorylation. It protects cells from stresses that cause ATP depletion by switching off ATP-consuming biosynthetic pathways.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Assay dilution & Images
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Direct ELISA, 0.1-0.5μg/ml
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of AMPK alpha 1 using anti-AMPK alpha 1 antibody (A00994-3).
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: human Jurkat 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-AMPK alpha 1 antigen affinity purified polyclonal antibody (Catalog # A00994-3) 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 AMPK alpha 1 at approximately 63KD. The expected band size for AMPK alpha 1 is at 63KD.
Click image to see more details
Figure 2. IF analysis of AMPK alpha 1 using anti-AMPK alpha 1 antibody (A00994-3).
AMPK alpha 1 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-AMPK alpha 1 Antibody (A00994-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 3. Flow Cytometry analysis of SiHa cells using anti-AMPK alpha 1 antibody (A00994-3).
Overlay histogram showing SiHa cells stained with A00994-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AMPK alpha 1 Antibody (A00994-3,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For PRKAA1 (Source: Uniprot.org, NCBI)
Gene Name
PRKAA1
Full Name
5'-AMP-activated protein kinase catalytic subunit alpha-1
Weight
Superfamily
protein kinase superfamily
Alternative Names
AMP -activate kinase alpha 1 subunit; AMP-activated protein kinase, catalytic, alpha-1,5'-AMP-activated protein kinase catalytic subunit alpha-1; AMPK alpha 1; AMPK alpha 1,5'-AMP-activated protein kinase, catalytic alpha-1 chain; AMPK subunit alpha-1; AMPK; AMPK1; AMPKa1; EC 2.7.11; EC 2.7.11.1; MGC33776; MGC57364; PRKAA1; protein kinase, AMP-activated, alpha 1 catalytic subunit PRKAA1 AMPK, AMPKa1 protein kinase AMP-activated catalytic subunit alpha 1 5-AMP-activated protein kinase catalytic subunit alpha-1|5-AMP-activated protein kinase, catalytic alpha-1 chain|ACACA kinase|AMP -activate kinase alpha 1 subunit|AMP-activated protein kinase, catalytic, alpha-1|AMPK alpha 1|AMPK subunit alpha-1|HMGCR kinase|acetyl-CoA carboxylase kinase|hydroxymethylglutaryl-CoA reductase kinase|protein kinase, AMP-activated, alpha 1 catalytic subunit|tau-protein kinase PRKAA1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on PRKAA1, check out the PRKAA1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for PRKAA1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-AMPK alpha 1/PRKAA1 Antibody Picoband™ (A00994-3)
Hello CJ!
A00994-3 has been cited in 2 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
The apolipoprotein A?I mimetic peptide, D?4F, restrains neointimal formation through heme oxygenase?1 up?regulation
Hou S, Zheng F, Li Y, Gao L, Zhang J. Int J Mol Sci. 2014 Aug 26;15(9):15026-43. Doi: 10.3390/Ijms150915026. The Protective Effect Of Glycyrrhizic Acid On Renal Tubular Epithelial Cell Injury Induced By High Glucose.
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Customer Q&As
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1 Customer Q&As for Anti-AMPK alpha 1/PRKAA1 Antibody Picoband™
Question
We are currently using anti-AMPK alpha 1/PRKAA1 antibody A00994-3 for rat tissue, and we are well pleased with the ELISA results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on dog tissues as well?
N. Wu
Verified customer
Asked: 2014-06-27
Answer
The anti-AMPK alpha 1/PRKAA1 antibody (A00994-3) has not been validated for cross reactivity specifically with dog tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2014-06-27