Recombinant Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] KO Tested (ab185141)

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Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18814-102] to GSK3 beta + GSK3 alpha
Suitable for: IHC-P, Flow Cyt, IP, ICC/IF, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102]
Description

Rabbit monoclonal [EPR18814-102] to GSK3 beta + GSK3 alpha
Host species

Rabbit
Tested applications

Suitable for: IHC-P, Flow Cyt, IP, ICC/IF, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment within Mouse GSK3 beta aa 200 to the C-terminus. The exact sequence is proprietary.
Database link: Q9WV60
Positive control
- WB: His-tagged mouse GSK3 beta (aa218-420) recombinant protein; His-tagged mouse GSK3 alpha (aa330-490) recombinant protein; Human fetal brain, fetal heart and fetal kidney lysates; NIH/3T3, HeLa, A549, RAW 264.7 and PC-12 whole cell lysates; Mouse and rat heart lysates. IHC-P: Mouse and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18814-102
Isotype

IgG
Research areas

Alternative Versions
- Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Our Abpromise guarantee covers the use of ab185141 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
IHC-P		1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This antibody is not recommended for IHC in human.
Flow Cyt		1/60.
IP		1/30.
ICC/IF		1/150.
WB		1/5000. Detects a band of approximately 47, 52 kDa (predicted molecular weight: 47, 52 kDa).

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
- Omim: 605004 Human
- Omim: 606784 Human
- SwissProt: P49840 Human
- SwissProt: P49841 Human
- SwissProt: Q2NL51 Mouse
- SwissProt: Q9WV60 Mouse
- SwissProt: P18265 Rat
- SwissProt: P18266 Rat
- Unigene: 445733 Human
- Unigene: 466828 Human
- Unigene: 294664 Mouse
- Unigene: 394930 Mouse
- Unigene: 10426 Rat
- Unigene: 36807 Rat
see all

Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: GSK3 alpha knockout HAP1 whole cell lysate (20 µg)
Lane 3: GSK3 beta whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)
Lane 5: Hek293 whole cell lysate (20 µg)

Lanes 1 - 5: Merged signal (red and green). Green - ab185141 observed at 47/52 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab185141 was shown to specifically react with GSK3 alpha and GSK3 beta in wild-type HAP1 cells as signal was lost in GSK3 alpha and GSK3 beta knockout cells. Wild-type and GSK3 alpha and GSK3 beta knockout samples were subjected to SDS-PAGE. ab185141 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling GSK3 beta + GSK3 alpha with ab185141 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse testis (PMID: 22792253). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/1000 dilution

Lane 1 : His-tagged mouse GSK3 beta (aa218-420) recombinant protein
Lane 2 : His-tagged mouse GSK3 alpha (aa330-490) recombinant protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 47, 52 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Developed using the ECL technique.

Predicted band size: 47, 52 kDa
Observed band size: 47,52 kDa
why is the actual band size different from the predicted?

Exposure time : Lane 1: 30 seconds; Lanes 2 and 3: 3 minutes.

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution

Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate
Lane 4 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 6 : Mouse heart lysate
Lane 7 : Rat heart lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 47, 52 kDa
Observed band size: 47,52 kDa why is the actual band size different from the predicted?

Exposure time : Lanes 1 and 2:10 seconds; Lane 3: 1 minute; Lanes 4 and 5: 5 seconds; Lane 6:3 seconds; Lane 7:10 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling GSK3 beta + GSK3 alpha with ab185141 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat testis (PMID: 22792253). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GSK3 beta + GSK3 alpha with ab185141 at 1/150 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling GSK3 beta + GSK3 alpha with ab185141 at 1/150 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow Cytometry - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cell line labeling GSK3 beta + GSK3 alpha with ab185141 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

GSK3 beta + GSK3 alpha was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab185141 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab185141 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: NIH/3T3 whole cell lysate 10 µg (Input).

Lane 2: ab185141 IP in NIH/3T3 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185141 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

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.

Click here to view the general protocols

Datasheet

Publishing research using ab185141? Please let us know so that we can cite the reference in this datasheet.

ab185141 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Recombinant Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

Target

Cellular localization

Database links

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (0)

Customer reviews and Q&As