重组Anti-Transcription factor AP-2-alpha抗体[EPR2688(2)] - BSA and Azide free (ab236043)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2688(2)] to Transcription factor AP-2-alpha - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Transcription factor AP-2-alpha抗体[EPR2688(2)] - BSA and Azide free
参阅全部 Transcription factor AP-2-alpha 一抗 -
描述
兔单克隆抗体[EPR2688(2)] to Transcription factor AP-2-alpha - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, ICC/IF, WB, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IP: HeLa whole cell lysate; Flow Cyt (intra): JAR cells; ICC/IF: JAR cells; IHC-P: Human breast carcinoma, and mouse and rat breast tissue; WB: HeLa, C6, Mouse skin and HAP1 cell lysates.
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常规说明
ab236043 is the carrier-free version of ab108311.
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: 7.2
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
EPR2688(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311)
- PE Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab305403)
- HRP Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab305405)
- Alexa Fluor® 488 Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab309663)
- Alexa Fluor® 647 Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab310026)
- Alexa Fluor® 594 Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab310404)
- Alexa Fluor® 555 Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab311931)
- Alexa Fluor® 568 Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab312402)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab236043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.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
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IP |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa. |
IP
Use at an assay dependent concentration. |
靶标
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功能
Sequence-specific DNA-binding protein that interacts with inducible viral and cellular enhancer elements to regulate transcription of selected genes. AP-2 factors bind to the consensus sequence 5'-GCCNNNGGC-3' and activate genes involved in a large spectrum of important biological functions including proper eye, face, body wall, limb and neural tube development. They also suppress a number of genes including MCAM/MUC18, C/EBP alpha and MYC. AP-2-alpha is the only AP-2 protein required for early morphogenesis of the lens vesicle. -
疾病相关
Defects in TFAP2A are the cause of branchiooculofacial syndrome (BOFS) [MIM:113620]; also known as branchial clefts with characteristic facies, growth retardation, imperforate nasolacrimal duct, and premature aging or lip pseudocleft-hemangiomatous branchial cyst syndrome. BOFS is a rare autosomal dominant cleft palate craniofacial disorder with variable expressivity. The major features include cutaneous anomalies, ocular anomalies, characteristic facial appearance (malformed pinnae, oral clefts), and, less commonly, renal and ectodermal (dental and hair) anomalies. -
序列相似性
Belongs to the AP-2 family. -
结构域
The WW-binding motif mediates interaction with WWOX. -
翻译后修饰
Sumoylated on Lys-10; which inhibits transcriptional activity. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 7020 Human
- Entrez Gene: 21418 Mouse
- Entrez Gene: 306862 Rat
- Omim: 107580 Human
- SwissProt: P05549 Human
- SwissProt: P34056 Mouse
- SwissProt: P58197 Rat
- Unigene: 519880 Human
see all -
别名
- Activating enhancer binding protein 2 alpha antibody
- Activating enhancer-binding protein 2-alpha antibody
- Activator protein 2 antibody
see all
图片
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All lanes : Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/10000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 3 : Mouse skin lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDaThis data was developed using ab108311, the same antibody clone in a different buffer formulation.
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This data was developed using ab108311, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of JAR (Human placenta choriocarcinoma epithelial cell) cells labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/50 dilution (3.4 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
This data was developed using ab108311, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat breast tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab108311, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse breast tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab108311, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab108311, the same antibody clone in a different buffer formulation.
Purified ab108311 at 1/20 dilution (0.5µg) immunoprecipitating Transcription factor AP-2-alpha in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab108311 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108311 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/10,000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 48 kDa -
This data was developed using ab108311, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of JAR (Human placenta choriocarcinoma epithelial cell) cells labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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All lanes : Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TFAP2A knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 48 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab108311).
Lanes 1- 2: Merged signal (red and green). Green - ab108311 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab108311 was shown to react with Transcription factor AP-2-alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265122 (knockout cell lysate ab257736) was used. Wild-type HeLa and TFAP2A knockout HeLa cell lysates were subjected to SDS-PAGE. ab108311 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 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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All lanes : Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TFAP2A (Transcription factor AP-2-alpha) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 48 kDaLanes 1 - 2: Merged signal (red and green). Green - ab108311 observed at 48 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab108311 was shown to recognize 0 in wild-type HAP1 cells as signal was lost at the expected MW in TFAP2A (AP2A) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TFAP2A (AP2A) knockout samples were subjected to SDS-PAGE. Ab108311 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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 1/20000 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 (ab108311).
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
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Datasheet download
Certificate of Compliance
文献 (0)
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