重组Anti-GSK3 beta抗体[Y174] - BSA and Azide free (ab183177)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ELISA, ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
概述
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产品名称
Anti-GSK3 beta抗体[Y174] - BSA and Azide free
参阅全部 GSK3 beta 一抗 -
描述
兔单克隆抗体[Y174] to GSK3 beta - BSA and Azide free -
宿主
Rabbit -
特异性
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.
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经测试应用
适用于: Flow Cyt (Intra), ELISA, ICC/IF, IHC-P, WBmore details -
种属反应性
与反应: Human, Recombinant fragment
预测可用于: Mouse -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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)
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常规说明
ab183177 is the carrier-free version of ab32391.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y174 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 647 Anti-GSK3 beta antibody [Y174] (ab196910)
- HRP Anti-GSK3 beta antibody [Y174] (ab196911)
- Alexa Fluor® 488 Anti-GSK3 beta antibody [Y174] (ab197236)
- Alexa Fluor® 594 Anti-GSK3 beta antibody [Y174] (ab201737)
- Alexa Fluor® 555 Anti-GSK3 beta antibody [Y174] (ab201738)
- PE Anti-GSK3 beta antibody [Y174] (ab210619)
- Anti-GSK3 beta antibody [Y174] (ab32391)
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Compatible Secondaries
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Conjugation kits
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Isotype control
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab183177于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ELISA |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 46 kDa.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ELISA
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 46 kDa. |
靶标
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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
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数据库链接
- Entrez Gene: 2932 Human
- Entrez Gene: 56637 Mouse
- Omim: 605004 Human
- SwissProt: P49841 Human
- SwissProt: Q9WV60 Mouse
- Unigene: 445733 Human
- Unigene: 394930 Mouse
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别名
- Glycogen Synthase Kinase 3 Beta antibody
- Glycogen synthase kinase-3 beta antibody
- GSK 3 beta antibody
see all
图片
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All lanes : HRP Anti-GSK3 beta antibody [Y174] (ab196911) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GSK3B knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDaab196911 was shown to recognize GSK3 beta in wild-type HAP1 cells as signal was lost at the expected MW in GSK3B knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GSK3B knockout samples were subjected to SDS-PAGE. Ab196911 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196911).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32391).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (7)
ab183177 被引用在 7 文献中.
- García-Montero C et al. Irregular Expression of Cellular Stress Response Markers in the Placenta of Women with Chronic Venous Disease. Antioxidants (Basel) 11:N/A (2022). PubMed: 36421463
- Wang WY et al. A Gene Expression Signature of Epithelial Tubulogenesis and a Role for ASPM in Pancreatic Tumor Progression. Gastroenterology 145:1110-20 (2013). WB ; Human . PubMed: 23896173
- Olsen AK et al. Regulation of APC and AXIN2 expression by intestinal tumor suppressor CDX2 in colon cancer cells. Carcinogenesis 34:1361-9 (2013). Human . PubMed: 23393221
- John JK et al. GSK3ß Inhibition Blocks Melanoma Cell/Host Interactions by Downregulating N-Cadherin Expression and Decreasing FAK Phosphorylation. J Invest Dermatol : (2012). PubMed: 22810307
- Ha GH et al. Tankyrase-1 function at telomeres and during mitosis is regulated by Polo-like kinase-1-mediated phosphorylation. Cell Death Differ 19:321-32 (2012). PubMed: 21818122
- Miyashita K et al. Potential therapeutic effect of glycogen synthase kinase 3beta inhibition against human glioblastoma. Clin Cancer Res 15:887-97 (2009). WB, IHC-P ; Human . PubMed: 19188159
- Juhlin CC et al. Loss of expression for the Wnt pathway components adenomatous polyposis coli and glycogen synthase kinase 3-beta in parathyroid carcinomas. Int J Oncol 34:481-92 (2009). IHC-P ; Human . PubMed: 19148484