Anti-GSK3 beta (phospho Y216) antibody (ab75745)

Rabbit polyclonal GSK3 beta (phospho Y216) antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 50 publication(s). Independently reviewed in 4 review(s).

Overview

  • Product name

    Anti-GSK3 beta (phospho Y216) antibody
    See all GSK3 beta primary antibodies
  • Description

    Rabbit polyclonal to GSK3 beta (phospho Y216)
  • Host species

    Rabbit
  • Specificity

    ab75745 detects endogenous levels of GSK3 beta only when phosphorylated at tyrosine 216. This antibody can also cross-react with GSK3 alpha subunit due to the consistent antigen-sequence of "GEPNVSYICSRYY".
  • Tested applications

    Suitable for: IHC-Fr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phosphopeptide derived from human GSK3 beta around the phosphorylation site of tyrosine 216 (V-S-YP-I-C).

  • Positive control

    • Human breast carcinoma tissue; 293 cell extracts, untreated or treated with insulin (40 µM, 30 min).

Properties

Applications

Our Abpromise guarantee covers the use of ab75745 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-Fr Use at an assay dependent concentration. PubMed: 22096577
ICC/IF 1/100 - 1/200.
WB 1/500 - 1/1000. Predicted molecular weight: 47 kDa.Can be blocked with GSK3 beta (phospho Y216) peptide (ab110532). This antibody can be used with studies using ab110532.
IHC-P 1/50 - 1/100.

Target

  • Function

    Participates in the Wnt signaling pathway. Implicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor JUN. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates MUC1 in breast cancer cells, and decreases the interaction of MUC1 with CTNNB1/beta-catenin. Phosphorylates CTNNB1/beta-catenin. Phosphorylates SNAI1. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization. Phosphorylates MACF1 and this phosphorylation inhibits the binding of MACF1 to microtubules which is critical for its role in bulge stem cell migration and skin wound repair.
  • Tissue specificity

    Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Phosphorylated by AKT1 and ILK1. Activated by phosphorylation at Tyr-216.
  • Cellular localization

    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.
  • Information by UniProt
  • Database links

  • Alternative names

    • Glycogen Synthase Kinase 3 Beta antibody
    • Glycogen synthase kinase-3 beta antibody
    • GSK 3 beta antibody
    • GSK-3 beta antibody
    • GSK3B antibody
    • GSK3B_HUMAN antibody
    • GSK3beta isoform antibody
    • Serine/threonine-protein kinase GSK3B antibody
    see all

Images

  • All lanes : Anti-GSK3 beta (phospho Y216) antibody (ab75745) at 1/500 dilution

    Lane 1 : 293 cell extracts, untreated
    Lane 2 : 293 cell extracts, treated with insulin (40µM, 30min)

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

  • ab75745, at a 1/50 dilution, staining GSK3 beta in paraffin embedded human breast carcinoma tissue by Immunohistochemistry, in the absence (left image) or presence (right image) of the immunizing phospho peptide.
  • ab75745 at 1/100 dilution staining GSK3 beta in human Hela cells by ICC/IF.

  • ab75745 staining GSK3 beta (phospho Y216) in medial entorhinal cortex of MPS IIIB mice by Immunohistochemistry (Frozen sections).

    Sections were cut sagittally and permeabilized in 1% Triton X-100 in PBS for 1 hour at ambient temperature. After washing with H2O2 in methanol, sections were incubated sequentially with 0.5% donkey serum, primary antibody and a biotin-labeled donkey secondary antibody in TBS + 2% BSA + 0.02% Tween 20. The signal was visualized with DAB and counterstained with hematoxylin.
  • All lanes : Anti-GSK3 beta (phospho Y216) antibody (ab75745) at 1/500 dilution

    Lane 1 : 293 cells untreated PMA
    Lane 2 : 293 cells treated PMA

    Predicted band size: 47 kDa

References

This product has been referenced in:

  • Chang PM  et al. Fucoidan?Fucoxanthin Ameliorated Cardiac Function via IRS1/GRB2/ SOS1, GSK3ß/CREB Pathways and Metabolic Pathways in Senescent Mice. Mar Drugs 17:N/A (2019). Read more (PubMed: 30669571) »
  • Sun S  et al. LncRNA UCA1 promotes cell proliferation, invasion and migration of laryngeal squamous cell carcinoma cells by activating Wnt/ß-catenin signaling pathway. Exp Ther Med 17:1182-1189 (2019). Read more (PubMed: 30679991) »
See all 67 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Western blot
Sample
Dictyostelium discoideum Cell lysate - whole cell (Dictyostelium cells)
Gel Running Conditions
Reduced Denaturing (15 %)
Loading amount
500000 cells
Specification
Dictyostelium cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 11 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (ARPE-19)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
125 ug/ml Cigarette Smoke Extract, 24 hr
Specification
ARPE-19
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Lei Wang

Verified customer

Submitted Jul 07 2017

Application
Western blot
Sample
Nematostella vectensis Tissue lysate - whole (Nematostella planulae 6dpf)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
50 µg
Specification
Nematostella planulae 6dpf
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jun 05 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain-White matter)
Permeabilization
Yes - 0.2%
Specification
Brain-White matter
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 16 2016

Application
Western blot
Sample
Rabbit Tissue lysate - whole (brain tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
brain tissue
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 03 2015

Answer

At Abcam we generally do not recommend re-use of the diluted antibody as signal quality may decrease. However if you were to do this, use of 0.05% sodium azide should be enough to keep any bacterial growth down. When using sodium azide in this solution please be sure to wash the blot thoroughly to remove any residual azide prior to the addition of any HRP conjugated secondary antibody as sodim azide will interfer with HRP detection systems.

Read More

Answer

There are generally a few reasons why doublets form in western blot. One is that the sample has undergone proteolysis, for this we recommend creating new samples using fresh buffers with new protease inhibitors includes and to keep the samples cold at all times during preparation. Another cause may be the existence of isoforms of the protein of interest. In review of GSK3 beta I do see that a second isoform exists which has only a 1.3kDa difference in size. As each antibody is directed against regions which are the same in both isoforms, it may be that your blot is showing the two isoforms present in your samples. We would recommend repeating the experiments using 293 cell extracts, treated with insulin (40µM, 30min) as a control alongside your samples and to use fresh buffers, protease inhibitors while keeping the samples cold.

Read More

Answer

Thanks for your enquiry.

Those two references posted on our online datasheet for ab75745 are the only publications we know of that have used this antibody.

I'm sorry I could not be of further assistance. Please contact us again if you have any further questions.

Read More

Answer

If your customer wants to detect the activated form of this enzyme, the ab75745 Anti-GSK3 beta (phospho Y216) antibodyis the right choice for her: According to SwissProt, it is activated by phosphorylation at Tyr-216.


The ab93926 Anti-GSK3 beta antibody [3D10]cannot differentiate between the activated and the inhibited form, so it should show the total level of the protein.


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