重组Anti-Annexin-6/ANXA6抗体[EPR19536] (ab201024)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19536] to Annexin-6/ANXA6
- Suitable for: WB, ICC/IF, IP, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Annexin-6/ANXA6抗体[EPR19536]
参阅全部 Annexin-6/ANXA6 一抗 -
描述
兔单克隆抗体[EPR19536] to Annexin-6/ANXA6 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IP, IHC-P, Flow Cyt (Intra)more details -
种属反应性
与反应: Human -
免疫原
This product was produced with the following immunogens:
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers. -
阳性对照
- WB: Human fetal liver, fetal heart, fetal kidney, spleen and skeletal muscle lysates; Jurkat, Raji, HeLa and HepG2 whole cell lysates. IHC-P: Human tonsil and pancreas tissues. ICC/IF: HepG2 and Raji cells. Flow Cyt (intra): HepG2 cells. IP: Human fetal liver whole cell lysate.
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常规说明
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19536 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab201024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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WB | (1) |
1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa).
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ICC/IF |
1/500.
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IP |
1/30.
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IHC-P |
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/600.
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说明 |
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WB
1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa). |
ICC/IF
1/500. |
IP
1/30. |
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/600. |
靶标
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功能
May associate with CD21. May regulate the release of Ca(2+) from intracellular stores. -
序列相似性
Belongs to the annexin family.
Contains 8 annexin repeats. -
结构域
A pair of annexin repeats may form one binding site for calcium and phospholipid. -
翻译后修饰
Phosphorylated in response to growth factor stimulation. -
细胞定位
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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数据库链接
- Entrez Gene: 309 Human
- Omim: 114070 Human
- SwissProt: P08133 Human
- Unigene: 412117 Human
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别名
- 67 kDa calelectrin antibody
- Annexin A6 antibody
- Annexin VI antibody
see all
图片
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ANXA6 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 76 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab201024 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.
ab201024 Anti-Annexin-6/ANXA6 antibody [EPR19536] was shown to specifically react with Annexin-6/ANXA6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265677 (knockout cell lysate ab257351) was used. Wild-type and Annexin-6/ANXA6 knockout samples were subjected to SDS-PAGE. ab201024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Annexin-6/ANXA6 with ab201024 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on Human tonsil is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Annexin-6/ANXA6 with ab201024 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm and membrane staining on the Human pancreas islet is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on HepG2 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab201024 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/600 dilution (red) compared with a a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling Annexin-6/ANXA6 with ab201024 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on Raji cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab201024 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024) at 1/1000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 76 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2 and 3: 5 seconds; Lane 4: 30 seconds.
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19536] (ab201024) at 1/5000 dilution
Lane 1 : Human skeletal muscle lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell 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: 76 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2,3,4 and 5: 30 seconds.
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Annexin-6/ANXA6 was immunoprecipitated from 0.35mg of Human fetal liver whole cell lysate with ab201024 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab201024 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Human fetal liver whole cell lysate,10µg (Input).
Lane 2: ab201024 IP in Human fetal liver whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab201024 in Human fetal liver whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
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