Recombinant Anti-PHKA1 antibody [EPR12118] (ab176338)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12118] to PHKA1
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PHKA1 antibody [EPR12118]
See all PHKA1 primary antibodies -
Description
Rabbit monoclonal [EPR12118] to PHKA1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide within Human PHKA1 aa 350-450 (Cysteine residue). The exact sequence is proprietary.
Database link: P46020 -
Positive control
- A431, Jurkat, HeLa, and Human skeletal muscle lysates. Paraffin-embedded Human skeletal muscle tissue. Permeabilized Hela cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12118 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab176338 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) |
1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 137 kDa.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Predicted molecular weight: 137 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Phosphorylase b kinase catalyzes the phosphorylation of serine in certain substrates, including troponin I. The alpha chain may bind calmodulin. -
Tissue specificity
Muscle specific. Isoform 1 is predominant in vastus lateralis muscle. Isoform 2 predominates slightly in heart, and it predominates clearly in the other tissues tested. -
Pathway
Glycan biosynthesis; glycogen metabolism. -
Involvement in disease
Glycogen storage disease 9D (GSD9D) [MIM:300559]: A metabolic disorder characterized by slowly progressive, predominantly distal muscle weakness and atrophy. Clinical features include exercise intolerance with early fatigability, pain, cramps and occasionally myoglobinuria. Note=The disease is caused by mutations affecting the gene represented in this entry. -
Sequence similarities
Belongs to the phosphorylase b kinase regulatory chain family. -
Post-translational
modificationsAlthough the final Cys may be farnesylated, the terminal tripeptide is probably not removed, and the C-terminus is not methylated. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 5255 Human
- Entrez Gene: 18679 Mouse
- Entrez Gene: 64561 Rat
- Omim: 311870 Human
- SwissProt: P46020 Human
- SwissProt: P18826 Mouse
- SwissProt: Q64649 Rat
- Unigene: 201379 Human
see all -
Alternative names
- 5330411D17 antibody
- 9830108K24Rik antibody
- kinase PHKA1 antibody
see all
Images
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All lanes : Anti-PHKA1 antibody [EPR12118] (ab176338) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PHKA1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-PHKA1 antibody [EPR12118] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab176338 was shown to bind specifically to PHKA1. A band was observed at 130 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in PHKA1 knockout cell line ab267337 (knockout cell lysate ab258111).
To generate this image, wild-type and PHKA1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-PHKA1 antibody [EPR12118] (ab176338) at 1/1000 dilution
Lane 1 : A431 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Human skeletal muscle lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 137 kDa -
Intracellular flow cytometric analysis of permeabilized Hela cells labeling PHKA1 with ab176338 at 1/10 dilution (red) or a rabbit IgG (negative) (green).
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Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling PHKA1 with ab176338 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab176338 has been referenced in 2 publications.
- Liu X et al. Metformin and Berberine suppress glycogenolysis by inhibiting glycogen phosphorylase and stabilizing the molecular structure of glycogen in db/db mice. Carbohydr Polym 243:116435 (2020). PubMed: 32532388
- Krag TO et al. Differential glucose metabolism in mice and humans affected by McArdle disease. Am J Physiol Regul Integr Comp Physiol 311:R307-14 (2016). PubMed: 27280431