Recombinant Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y365] to AMPK alpha 1 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free
See all AMPK alpha 1 primary antibodies -
Description
Rabbit monoclonal [Y365] to AMPK alpha 1 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody is specific for human AMPK alpha 1. This antibody shows low affinity on mouse and rat samples.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HepG2, C6, NIH/3T3 and MCF-7 cell lysate. Mouse liver, brain, retina, and skeletal muscle tissue lysates. IHC-P: Human cervical carcinoma and lung carcinoma tissues. ICC/IF: MCF-7 cells. Flow Cyt (intra): HeLa cells. IP: HeLa cell lysate.
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General notes
ab210714 is the carrier-free version of ab32047.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y365 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- PE Anti-AMPK alpha 1 antibody [Y365] (ab303053)
- APC Anti-AMPK alpha 1 antibody [Y365] (ab303054)
- HRP Anti-AMPK alpha 1 antibody [Y365] (ab303055)
- Alexa Fluor® 488 Anti-AMPK alpha 1 antibody [Y365] (ab309679)
- Alexa Fluor® 647 Anti-AMPK alpha 1 antibody [Y365] (ab310041)
- Alexa Fluor® 594 Anti-AMPK alpha 1 antibody [Y365] (ab310429)
- Alexa Fluor® 555 Anti-AMPK alpha 1 antibody [Y365] (ab311955)
- Alexa Fluor® 568 Anti-AMPK alpha 1 antibody [Y365] (ab312428)
- Anti-AMPK alpha 1 antibody [Y365] (ab32047)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab210714 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 63 kDa.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 63 kDa. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Responsible for the regulation of fatty acid synthesis by phosphorylation of acetyl-CoA carboxylase. It also regulates cholesterol synthesis via phosphorylation and inactivation of hormone-sensitive lipase and hydroxymethylglutaryl-CoA reductase. Appears to act as a metabolic stress-sensing protein kinase switching off biosynthetic pathways when cellular ATP levels are depleted and when 5'-AMP rises in response to fuel limitation and/or hypoxia. This is a catalytic subunit. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Contains 1 protein kinase domain. - Information by UniProt
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Database links
- Entrez Gene: 5562 Human
- Entrez Gene: 105787 Mouse
- Entrez Gene: 65248 Rat
- Omim: 602739 Human
- SwissProt: Q13131 Human
- SwissProt: Q5EG47 Mouse
- SwissProt: P54645 Rat
- Unigene: 43322 Human
see all -
Alternative names
- 5 AMP activated protein kinase alpha 1catalytic subunit antibody
- 5 AMP activated protein kinase catalytic alpha 1 chain antibody
- 5' AMP activated protein kinase catalytic subunit alpha 1 antibody
see all
Images
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All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/1000 dilution
Lane 1 : Wild-type RAW 264.7 cell lysate
Lane 2 : PRKAA1 knockout RAW 264.7 cell lysate
Lane 3 : Mouse Liver cell lysate
Lane 4 : Neuro2A cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDaFalse colour image of Western blot: Anti-AMPK alpha 1 antibody [Y365] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32047 was shown to bind specifically to AMPK alpha 1. A band was observed at 64 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PRKAA1 knockout cell line ab280055 (knockout cell lysate ab280114). To generate this image, wild-type and PRKAA1 knockout RAW 264.7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye$®$ 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye$®$ 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling AMPK alpha 1 with purified ab32047 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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Immunocytochemistry/Immunofluorescence analysis of MCF7 cells labelling AMPK alpha 1 with purified ab32047 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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Intracellular Flow Cytometry analysis of HeLa cells labelling AMPK alpha 1 with purified ab32047 at 1/150 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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ab32047 (purified) at 1/40 immunoprecipitating AMPK alpha 1 in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab32047 + HeLa whole cell lysate (10µg).
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32047 in HeLa whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/500 dilution
Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Mouse retina tissue lysate
Lane 7 : Mouse skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 63 kDa
Observed band size: 63 kDa
Additional bands at: 40 kDa (possible non-specific binding)
Exposure time: 3 minutesBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
This antibody shows low affinity on mouse samples. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/20000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 6 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
Lane 7 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 63 kDa
Observed band size: 63 kDa
Exposure time: 180 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
This antibody shows low affinity on mouse and rat samples.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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Overlay histogram showing HeLa cells stained with unpurified ab32047 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab32047, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analaysis of human cervical carcinoma tissue labelling AMPK alpha 1 with unpurified ab32047 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab210714 has not yet been referenced specifically in any publications.