Anti-FOXA2抗体(ab5074)

Reviews (1)Q&A (4)References (11)

Key features and details

  • Goat polyclonal to FOXA2
  • Suitable for: IHC-P, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-FOXA2抗体
    参阅全部 FOXA2 一抗

  • 描述

    山羊多克隆抗体to FOXA2

  • 宿主

    Goat

  • 特异性

    This antibody is expected to recognise both reported isoforms.

  • 经测试应用

    适用于: IHC-P, WBmore details

  • 种属反应性

    与反应: Rat, Human
    预测可用于: Chicken, Xenopus laevis

  • 免疫原

    Synthetic peptide: GVYSRPIMNSS, corresponding to amino acids 447-457 of Human FOXA 2.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

性能

应用

Our Abpromise guarantee covers the use of ab5074 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P Use a concentration of 4 - 8 µg/ml.
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 48 kDa).

A 1 hour primary incubation is recommended for this product. A nuclear extraction procedure may be required to concentrate the protein in samples.

靶标

  • 功能

    Transcription factor that is involved in embryonic development, establishment of tissue-specific gene expression and regulation of gene expression in differentiated tissues. Is thought to act as a 'pioneer' factor opening the compacted chromatin for other proteins through interactions with nucleosomal core histones and thereby replacing linker histones at target enhancer and/or promoter sites. Binds DNA with the consensus sequence 5'-[AC]A[AT]T[AG]TT[GT][AG][CT]T[CT]-3' (By similarity). In embryonic development is required for notochord formation. Involved in the development of multiple endoderm-derived organ systems such as the liver, pancreas and lungs; FOXA1 and FOXA2 seem to have at least in part redundant roles. Originally discribed as a transcription activator for a number of liver genes such as AFP, albumin, tyrosine aminotransferase, PEPCK, etc. Interacts with the cis-acting regulatory regions of these genes. Involved in glucose homeostasis; regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis. Involved in regulation of fat metabolism. Binds to fibrinogen beta promoter and is involved in IL6-induced fibrinogen beta transcriptional activation.

  • 序列相似性

    Contains 1 fork-head DNA-binding domain.

  • 翻译后修饰

    Phosphorylation on Thr-156 abolishes binding to target promoters and subsequent transcription activation upon insulin stimulation.

  • 细胞定位

    Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm in a CRM1-dependent manner and in response to insulin signaling via AKT1 is exported from the nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Forkhead box A2 antibody
      • Forkhead box protein A2 antibody
      • FOX A2 antibody
      • foxa2 antibody
      • FOXA2_HUMAN antibody
      • Hepatic nuclear factor 3 beta antibody
      • Hepatocyte nuclear factor 3 antibody
      • Hepatocyte nuclear factor 3 beta antibody
      • Hepatocyte nuclear factor 3-beta antibody
      • HNF 3B antibody
      • HNF-3-beta antibody
      • HNF-3B antibody
      • HNF3B antibody
      • MGC19807 antibody
      • TCF 3B antibody
      • TCF-3B antibody
      • TCF3B antibody
      • Transcription factor 3B antibody
      see all

    图片

    • Western blot - Anti-FOXA2 antibody (ab5074)
      Western blot - Anti-FOXA2 antibody (ab5074)
      Anti-FOXA2 antibody (ab5074) at 1 µg/ml + Rat Lung tissue lysate at 35 µg

      Predicted band size: 48 kDa
      Observed band size: 60 kDa
      why is the actual band size different from the predicted?



      Primary incubation was 1 hour. Detected by chemiluminescence.

    • Western blot - Anti-FOXA2 antibody (ab5074)
      Western blot - Anti-FOXA2 antibody (ab5074)
      Anti-FOXA2 antibody (ab5074) at 1 µg/ml + Human Liver lysate at 1 µg/ml

      Predicted band size: 48 kDa
      Observed band size: 60 kDa why is the actual band size different from the predicted?



      Primary incubation was 1 hour. Detected by chemiluminescence.

    文献 (11)

    发表研究结果有使用 ab5074？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab5074 被引用在 11 文献中.

    • Soda M  et al. Repeated human deciduous tooth-derived dental pulp cell reprogramming factor transfection yields multipotent intermediate cells with enhanced iPS cell formation capability. Sci Rep 9:1490 (2019). PubMed: 30728386
    • Shen J  et al. Loss of FoxA2 accelerates neoplastic changes in the intrahepatic bile duct partly via the MAPK signaling pathway. Aging (Albany NY) 11:9280-9294 (2019). PubMed: 31689237
    • Bhanu NV  et al. Histone modification profiling reveals differential signatures associated with human embryonic stem cell self-renewal and differentiation. Proteomics 16:448-58 (2016). Flow Cyt ; Human . PubMed: 26631989
    • Saitoh I  et al. Choice of Feeders Is Important When First Establishing iPSCs Derived From Primarily Cultured Human Deciduous Tooth Dental Pulp Cells. Cell Med 8:9-23 (2015). PubMed: 26858904
    • Maier JA  et al. Foxa1 and Foxa2 are required for formation of the intervertebral discs. PLoS One 8:e55528 (2013). IHC . PubMed: 23383217
    View all Publications for this product

    客户评价及客户问答

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    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-FOXA2 antibody

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    60 µg
    Gel Running Conditions
    Reduced Denaturing (8)
    Sample
    Xenopus laevis Tissue lysate - whole (whole embryo)
    Specification
    whole embryo
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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    Abcam user community

    Verified customer

    提交于 Apr 10 2015

    Question

    Welche Isoformen erkennen Ihre FoxA2 AK?

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    Answer

    Vielen Dank für Ihren Anruf.

    Wie versprochen habe ich alle unsere FOXA2-Antikörper darauf geprüft, welche Isoformen von FOXA2 sie erkennen. Leider sind alle Immunogene in Regionen lokalisiert, die in beiden Isoformen vorkommen (Details siehe jeweiliges Datenblatt), und daher detektieren die Antikörper immer beide Isoformen. Wir haben also leider keinen Antikörper erhältlich, der zwischen den Isoformen unterscheiden kann.

    Es tut mir leid, dass ich keine besseren Nachrichten für Sie habe. Ich hoffe, diese Informationen helfen Ihnen dennoch weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM no signal SAMPLE mouse liver whole cell extract PRIMARY ANTIBODY 1:1000 dilution in 5% BSA in TBST, 4C overnight wash 3x5min with TBST DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED positive controls: mouse liver, mouse hepatocyte ANTIBODY STORAGE CONDITIONS stored in -20C in 20uL aliquots SAMPLE PREPARATION homogenize 100mg liver in 3mL SDS lysis buffer containing protease (aprotinin, leupeptin, pepstatin, NaF, PMSF, sodium orthovanadate, DTT) AMOUNT OF PROTEIN LOADED 100ug ELECTROPHORESIS/GEL CONDITIONS 10% acrylamide TRANSFER AND BLOCKING CONDITIONS block in 5% milk in TBST 1hr at room temp SECONDARY ANTIBODY rabbit anti-goat IgG HRP 1:2500 dilution RT x 2hr wash 3x5min with TBST HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary and secondary antibody concentration primary antibody diluent amount protein loaded (up to 200ug)

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    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. This antibody is yet to be tested in mouse cells. However, I would predict a favourable result given that the immunogen used to raise this antibody has 100% identity with the mouse homolog. You are largely using an approach that I would recommend. However, we have received a good Abreview from a customer using this antibody; albeit against human cells. The customer applied this antibody with favourable results using a dilution of 1/500. Therefore I would like to recommend that you you try applying this antibody at a reduced dilution. Given that you are preparing a tissue lysate I would also like to recommend that you try probing your blot using an antibody against a housekeeping protein such as GAPDH or beta actin. This will confirm the integrity of this lysate preparation as I would certainly expect detection of this protein in liver cells given the mass of protein that you have been loading. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 45090181 DESCRIPTION OF THE PROBLEM No bands SAMPLE Differentiated ES cell lysates, mouse and human. PRIMARY ANTIBODY Abcam goat poly anti-Foxa2, ab5074., dilution 0.5-1ugm/ml, at 4C o/n and RT2 hrs. Washed at RT 10 mins each time X 2. DETECTION METHOD ECLPlus POSITIVE AND NEGATIVE CONTROLS USED HepG2 cell lysates as positive. Undifferentiated ES cell lysate as negative. ANTIBODY STORAGE CONDITIONS Aliquoted and stored at -20?C SAMPLE PREPARATION Detergent containing buffer plus all the PI inhibitors for cell lysis. For gel loading-- usual gel loading buffer containing glycerol and SDS. Boiled sample at 100C for 5 mins. AMOUNT OF PROTEIN LOADED 50-100 ugm per lane ELECTROPHORESIS/GEL CONDITIONS Reducing, 10-12% gels. TRANSFER AND BLOCKING CONDITIONS 1-1.5 hrs using Tris-glycine buffer. Blocking agent 5% non-fat milk. SECONDARY ANTIBODY [another company] donkey anti-goat 1:2000, RT 45 mins. Wash at RT 10 min each X 2. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Incubation temp for primary Ab, time of incubation, concentration/dilution of primay Ab and secondary Ab.

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    Answer

    I'm sorry to hear you are having a problem with ab5074. This antibody has not been tested in HepG2 cells however it has been tested in human liver lysate and as HepG2 cells are liver derived I would suspect they express good levels of FOxa2. However, they may still express lower levels that liver tissue therefore I would like to recommend to do a nuclear extraction of the protein and to run the positive control of liver lysate along your samples. I would also like to recommend to use 5% BSA in TBST to block the membrane and to try blocking for 1 hour only (and incubating the antibody in TBST only). Finally, can I please confirm that you have checked the transfer of protein is adequate and that the secondary antibody used works with other goat primary antibodies (i.e. that it is not damaged). Please let me know if this helps and do not hesitate to contact me again if you still experience problems with those modifications.

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    Question

    Is a nuclear extraction prep sufficient to demonstrate Foxa2 activation or do I have to do a total cell lysate prep?

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    Answer

    Thank you for your enquiry and your interest in our products. Ab5074 recognizes Foxa2 which is a transcription activator for a number of liver genes. As the legend of the Western image on the on-line datasheet shows this antibody has been tested for this application with human liver (whole) tissue lysate. Since the expression levels of Foxa2 in liver cells are very high, normally you do not need to prepare nuclear extract. In case, you would plan to use cell lysate from different tissue or cells than perhaps it would be advisable to prepare nuclear extract. We would strongly suggest running human liver lysate as good positive control along with the samples to see if the detection system and the experimental conditions are fine. We hope this information is useful for you.

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