重组Anti-GSK3 beta + GSK3 alpha抗体[EPR18814-102] - BSA and Azide free (ab226169)
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 - BSA and Azide free
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
-
产品名称
Anti-GSK3 beta + GSK3 alpha抗体[EPR18814-102] - BSA and Azide free -
描述
兔单克隆抗体[EPR18814-102] to GSK3 beta + GSK3 alpha - BSA and Azide free -
宿主
Rabbit -
特异性
Unsuitable for human IHC-P.
-
经测试应用
适用于: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- IHC-P: Mouse testis tissue.
-
常规说明
ab226169 is the carrier-free version of ab185141.
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
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18814-102 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab226169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 |
---|---|---|
WB |
Use at an assay dependent concentration. Detects a band of approximately 47, 52 kDa (predicted molecular weight: 47, 52 kDa).
|
|
IHC-P |
Use at an assay dependent concentration. 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. |
|
ICC/IF |
Use at an assay dependent concentration.
|
|
IP |
Use at an assay dependent concentration.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
说明 |
---|
WB
Use at an assay dependent concentration. Detects a band of approximately 47, 52 kDa (predicted molecular weight: 47, 52 kDa). |
IHC-P
Use at an assay dependent concentration. 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. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
靶标
-
细胞定位
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. -
数据库链接
- 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
see all
图片
-
All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GSK3 alpha knockout HAP1 whole cell lysate
Lane 3 : GSK3 beta whole cell lysate
Lane 4 : HeLa whole cell lysate
Lane 5 : Hek293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 47, 52 kDaLanes 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).
-
Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
实验方案
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.
数据表及文件
-
Datasheet download
Certificate of Compliance
文献 (1)
ab226169 被引用在 1 文献中.
- Sheng H et al. Combined Pharmacotherapy with Alendronate and Desferoxamine Regulate the Bone Resorption and Bone Regeneration for Preventing Glucocorticoids-Induced Osteonecrosis of the Femoral Head. Biomed Res Int 2020:3120458 (2020). PubMed: 33029500