Recombinant Anti-GSK3 beta (phospho Y216) + GSK3 alpha (phospho Y279) antibody [EPR933Y] - BSA and Azide free (ab239862)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR933Y] to GSK3 beta (phospho Y216) + GSK3 alpha (phospho Y279) - BSA and Azide free
- Suitable for: IHC-P, Dot blot, IP, WB
- Reacts with: Mouse, Rat, Human, Zebrafish
Related conjugates and formulations
Overview
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Product name
Anti-GSK3 beta (phospho Y216) + GSK3 alpha (phospho Y279) antibody [EPR933Y] - BSA and Azide free -
Description
Rabbit monoclonal [EPR933Y] to GSK3 beta (phospho Y216) + GSK3 alpha (phospho Y279) - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody detects both GSK alpha phosphorylated on tyrosine279 and GSK3 beta phosphorylated on tyrosine 216.
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Tested applications
Suitable for: IHC-P, Dot blot, IP, WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human, Zebrafish -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC: Human thyroid carcinoma, human ovarian carcinoma.
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General notes
ab239862 is the carrier-free version of ab68476.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR933Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239862 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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IHC-P |
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This antibody may not be suitable for IHC with mouse, rat or zebrafish samples. |
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Dot blot |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 47-52 kDa (predicted molecular weight: 47 kDa).
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Notes |
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IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. This antibody may not be suitable for IHC with mouse, rat or zebrafish samples. |
Dot blot
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 47-52 kDa (predicted molecular weight: 47 kDa). |
Target
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Cellular localization
GSK3 beta: Cytoplasm. Nucleus. Cell membrane. The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosophorylated form to the cell membrane. -
Database links
- Entrez Gene: 2931 Human
- Entrez Gene: 2932 Human
- Entrez Gene: 56637 Mouse
- Entrez Gene: 606496 Mouse
- Entrez Gene: 50686 Rat
- Entrez Gene: 84027 Rat
- Entrez Gene: 30654 Zebrafish
- Entrez Gene: 30664 Zebrafish
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling GSK3 (alpha + beta) with purified ab68476 at 1/2000. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. ab209101, a Rabbit specific IHC polymer detection kit HRP/DAB was used. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Positive staining on human thyroid carcinoma without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B).
The section was incubated with ab68476 for 30 minutes at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68476).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling GSK3 (alpha + beta) with purified ab68476 at 1/2000. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. ab209101, a Rabbit specific IHC polymer detection kit HRP/DAB was used. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Positive staining on human ovarian carcinoma without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B).
The section was incubated with ab68476 for 30 minutes at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68476).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling GSK3 (alpha + beta) with Purified ab68476 at 1:50 dilution (5.3 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68476).
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GSK3 (alpha + beta) (phospho Y216 + Y279) was immunoprecipitated from 10μg HeLa (human cervix adenocarcinoma) whole cell lysate with ab68476 at 1/50 dilution (2μg in 0.35mg lysates). Western blot was performed from the immunoprecipitate using ab68476 at 1/200 dilution (9 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1 (Input): HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysate 10μg
Lane 2 (+): HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab68476 in HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysateExposure Time: 30 seconds
Blocking and 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 (ab68476).
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Dot Blot analysis of Lane 1: Human GSK3 (alpha + beta) (pY216 + pY279) phospho peptide and Lane 2: Human GSK3 (alpha + beta) non-phospho peptide labeling GSK3 (alpha + beta) (phospho Y216 + Y279) with ab68476 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68476).
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Human brain tissue stained with unpurified ab68476 at 1/100 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68476).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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Datasheet download
Certificate of Compliance
References (0)
ab239862 has not yet been referenced specifically in any publications.