Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y174] to GSK3 beta
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, ELISA
- Knockout validated
- Reacts with: Mouse, Human
参阅全部 GSK3 beta 一抗
描述兔单克隆抗体[Y174] to GSK3 beta
This antibody is specific for human GSK3 beta. It may also detect the splice isoform 2 based on sequence homology.
The immunogen used for this antibody is GSK3 beta phospho S9. This antibody shows partially phospho specificity to phospho S9 under certain conditions, for example, under low peptide concentration in ELISA assay, it has dominant reactivity with phospho S9 peptide.
经测试应用适用于: ICC/IF, WB, IHC-P, Flow Cyt, ELISAmore details
种属反应性与反应: Mouse, Human
Synthetic peptide within Human GSK3 beta aa 350-450 (N terminal). The exact sequence is proprietary.
- A431 cell lysate. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145. IHC-P: Human breast adenocarcinoma FFPE tissue sections. ICC/IF: HeLa whole cell lysate (ab150035)
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.
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.
存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度Protein A purified
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Integration of energy
- Anti-GSK3 beta antibody [Y174] - BSA and Azide free (ab183177)
- Anti-GSK3 beta antibody [Y174] (Alexa Fluor® 647) (ab196910)
- Anti-GSK3 beta antibody [Y174] (HRP) (ab196911)
- Anti-GSK3 beta antibody [Y174] (Alexa Fluor® 488) (ab197236)
- Anti-GSK3 beta antibody [Y174] (Alexa Fluor® 594) (ab201737)
- Anti-GSK3 beta antibody [Y174] (Alexa Fluor® 555) (ab201738)
- Anti-GSK3 beta antibody [Y174] (PE) (ab210619)
Our Abpromise guarantee covers the use of ab32391 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/100 - 1/500.
|WB||1/5000 - 1/10000. Predicted molecular weight: 46 kDa.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ELISA||Use at an assay dependent concentration.
Unit Type: 0 - 1000 ng/ml
功能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.
组织特异性Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney.
序列相似性Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
Contains 1 protein kinase domain.
翻译后修饰Phosphorylated by AKT1 and ILK1. Activated by phosphorylation at Tyr-216.
细胞定位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
- Glycogen Synthase Kinase 3 Beta antibody
- Glycogen synthase kinase-3 beta antibody
- GSK 3 beta antibody
Lane 1 & 3: Wild type HAP1 whole cell lysate (20 µg)
Lane 2 & 4: GSK3B knockout HAP1 whole cell lysate (20 µg)
Lanes 1 - 4: Green - ab32391 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32391 was shown to recognize GSK3B in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when GSK3B knockout samples were examined. Wild-type and GSK3B knockout samples were subjected to SDS-PAGE. Ab32391 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical analysis of paraffin-embedded human breast tissue labelled with untreated ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (phospho) (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling GSK3 beta with purified ab32391 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (ab150120).
For negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.
ELISA of GSK3 beta (phospho S9) peptide and GSK3 beta non-phospho peptide at 10 ng/ml. Detected with ab32391 at 0~1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 was used as a secondary antibody.
ELISA of GSK3 beta (phospho S9) peptide and GSK3 beta non-phospho peptide at 1000 ng/ml. Detected with ab32391 at 0~1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 was used as a secondary antibody.
Overlay histogram showing HeLa cells stained with ab32391 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32391, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Anti-GSK3 beta antibody [Y174] (ab32391) at 1/10000 dilution + A431 cell lysate.
Predicted band size: 46 kDa
Observed band size: 46 kDa
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelled with untreated ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (phospho) (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
IHC image of GSK3 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab32391, 1/200 diution, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab32391 stained DU145 cells. The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1 %BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS-Tween for 1 hour to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32391 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
All lanes : Anti-GSK3 beta antibody [Y174] (ab32391) at 1/2500 dilution
Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : SW480 (human colorectal adenocarcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Donkey polyclonal IRDye 800CW at 1/15000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Additional bands at: 37 kDa. We are unsure as to the identity of these extra bands.
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.
ab32391 被引用在 88 文献中.
- Jiang N et al. HIF-1?-regulated miR-1275 maintains stem cell-like phenotypes and promotes the progression of LUAD by simultaneously activating Wnt/ß-catenin and Notch signaling. Theranostics 10:2553-2570 (2020). PubMed: 32194819
- Cheng J et al. Catalpol Promotes the Proliferation and Differentiation of Osteoblasts Induced by High Glucose by Inhibiting KDM7A. Diabetes Metab Syndr Obes 13:705-712 (2020). PubMed: 32214833
- Li Q et al. Polyphyllin I attenuates pressure over-load induced cardiac hypertrophy via inhibition of Wnt/ß-catenin signaling pathway. Life Sci 252:117624 (2020). PubMed: 32259602
- Zhang J et al. N1-Methylnicotinamide Improves Hepatic Insulin Sensitivity via Activation of SIRT1 and Inhibition of FOXO1 Acetylation. J Diabetes Res 2020:1080152 (2020). PubMed: 32280711
- Ou B et al. A positive feedback loop of ß-catenin/CCR2 axis promotes regorafenib resistance in colorectal cancer. Cell Death Dis 10:643 (2019). PubMed: 31501414