Recombinant Anti-GAK antibody [EPR25327-39] (ab305101)
![Western blot - Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-494110-anti-gak-antibody-epr25327-39-western-blot-1.jpg "Western blot - Anti-GAK antibody [EPR25327-39] (ab305101)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25327-39] to GAK
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-GAK antibody [EPR25327-39]
See all GAK primary antibodies -
Description
Rabbit monoclonal [EPR25327-39] to GAK -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt (Intra),ICC/IF or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Whole cell lysates: HEK-293T (human embryonic kidney epithelial cell), GAK knockout HeLa, Daudi (human Burkitt's lymphoma lymphoblast), Raji (human Burkitt's lymphoma B lymphocyte): Tissue lysates: Human testis and kidney. IHC-P: Human testis, breast carcinoma. HEK-293T wild type and GAK knockout.
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR25327-39 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab305101 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa).
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| IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa). |
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IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Associates with cyclin G and CDK5. Seems to act as an auxilin homolog that is involved in the uncoating of clathrin-coated vesicles by Hsc70 in non-neuronal cells. Expression oscillates slightly during the cell cycle, peaking at G1. -
Tissue specificity
Ubiquitous. Highest in testis. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
Contains 1 C2 tensin-type domain.
Contains 1 J domain.
Contains 1 phosphatase tensin-type domain.
Contains 1 protein kinase domain. -
Cellular localization
Cytoplasm > perinuclear region. Golgi apparatus > trans-Golgi network. Cell junction > focal adhesion. Localizes to the perinuclear area and to the trans-Golgi network. Also seen on the plasma membrane, probably at focals adhesions. - Information by UniProt
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Database links
- Entrez Gene: 2580 Human
- Omim: 602052 Human
- SwissProt: O14976 Human
- Unigene: 369607 Human
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Alternative names
- Auxilin 2 antibody
- Cyclin G associated kinase antibody
- Cyclin-G-associated kinase antibody
see all
Images
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Western blot - Anti-GAK antibody [EPR25327-39] (ab305101)All lanes : Anti-GAK antibody [EPR25327-39] (ab305101) at 1/1000 dilution
Lane 1 : Wild-type 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 2 : GAK knockout HeLa whole cell lysate
Lane 3 : Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), 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: 143 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?
Exposure time: 48 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, ab305101 was shown to bind specifically to GAK. A band was observed at 160 kDa in wild-type 293T cell lysates with no signal observed at this size in GAK knockout cell line ab266417 (knockout cell lysate ab263210).
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![Western blot - Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-494109-anti-gak-antibody-epr25327-39-western-blot-2.jpg)
Western blot - Anti-GAK antibody [EPR25327-39] (ab305101)All lanes : Anti-GAK antibody [EPR25327-39] (ab305101) at 1/1000 dilution
Lane 1 : Human testis tissue lysate
Lane 2 : Human kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 143 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?
Exposure time: 92 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-1-anti-gak-antibody-epr25327-39-immunohistochemistry-human-testis.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)Immunohistochemical analysis of paraffin-embedded human testis tissue labeling GAK with ab305101 at 1/50 dilution (9.9 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human testis is observed (PMID: 9299234). The section was incubated with ab305101 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-2-anti-gak-antibody-epr25327-39-immunohistochemistry-human-breast-carcinoma.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling GAK with ab305101 at 1/50 dilution (9.9 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human breast carcinoma is observed (PMID: 16161052). The section was incubated with ab305101 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-3-anti-gak-antibody-epr25327-39-immunohistochemistry-hek293.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAK antibody [EPR25327-39] (ab305101)Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK293 tissue labeling GAK with ab305101 at 1/50 dilution (9.9 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on (A) Wild-type HEK293T cell pellet is observed. No staining is observed on (B) GAK knockout HEK293T cell pellet. The section was incubated with ab305101 at 4°C overnight. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
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![Anti-GAK antibody [EPR25327-39] (ab305101)](https://www.abcam.com/ps/products/305/ab305101/Images/ab305101-4-Benefits-of-Recombinant-antibodies.jpg)
Anti-GAK antibody [EPR25327-39] (ab305101)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab305101 has not yet been referenced specifically in any publications.