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使用敲除细胞株进行验证

Anti-FOXA1抗体[1B1] (ab55178)

Reviews (1)Q&A (3)References (14)
Western blot - Anti-FOXA1 antibody [1B1] (ab55178)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [1B1] (ab55178)
  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [1B1] (ab55178)
  • Western blot - Anti-FOXA1 antibody [1B1] (ab55178)
  • Flow Cytometry - Anti-FOXA1 antibody [1B1] (ab55178)

Key features and details

  • Mouse monoclonal [1B1] to FOXA1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2a

选择批间可重复性更高的重组抗体

Product image
Anti-FOXA1 antibody [EPR10881] (ab170933)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术，可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-FOXA1抗体[1B1]
    参阅全部 FOXA1 一抗

  • 描述

    小鼠单克隆抗体[1B1] to FOXA1

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, IHC-P, ICC/IF, Flow Cytmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant fragment corresponding to Human FOXA1 aa 367-472.
    Database link: P55317

  • 阳性对照

    • WB: HepG2 and HeLa cell lysates.

  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 24/1/19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab55178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. Predicted molecular weight: 49 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
ICC/IF (1)
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
说明
WB
Use at an assay dependent concentration. Predicted molecular weight: 49 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

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). Proposed to play a role in translating the epigenetic signatures into cell type-specific enhancer-driven transcriptional programs. Its differential recruitment to chromatin is dependent on distribution of histone H3 methylated at 'Lys-5' (H3K4me2) in estrogen-regulated genes. Involved in the development of multiple endoderm-derived organ systems such as liver, pancreas, lung and prostate; FOXA1 and FOXA2 seem to have at least in part redundant roles (By similarity). Modulates the transcriptional activity of nuclear hormone receptors. Is involved in ESR1-mediated transcription; required for ESR1 binding to the NKX2-1 promoter in breast cancer cells; binds to the RPRM promter and is required for the estrogen-induced repression of RPRM. Involved in regulation of apoptosis by inhibiting the expression of BCL2. Involved in cell cycle regulation by activating expression of CDKN1B, alone or in conjunction with BRCA1. 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.

  • 组织特异性

    Highly expressed in prostate and ESR1-positive breast tumors. Overexpressed in esophageal and lung adenocarcinomas.

  • 序列相似性

    Contains 1 fork-head DNA-binding domain.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • forkhead box A1 antibody
      • Forkhead box protein A1 antibody
      • FOX A1 antibody
      • FOXA1 antibody
      • FOXA1_HUMAN antibody
      • hepatocyte nuclear factor 3 alpha antibody
      • Hepatocyte nuclear factor 3-alpha antibody
      • HNF 3A antibody
      • HNF-3-alpha antibody
      • HNF-3A antibody
      • HNF3A antibody
      • MGC33105 antibody
      • TCF 3A antibody
      • TCF-3A antibody
      • TCF3A antibody
      • Transcription factor 3A antibody
      see all

    图片

    • Western blot - Anti-FOXA1 antibody [1B1] (ab55178)
      Western blot - Anti-FOXA1 antibody [1B1] (ab55178)
      All lanes : Anti-FOXA1 antibody [1B1] (ab55178) at 1/500 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : FOXA1 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 49 kDa
      Observed band size: 49 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab55178 observed at 49 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

       ab55178 was shown to react with FOXA1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261823 (knockout cell lysate ab256920) was used. Wild-type HeLa and FOXA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab55178 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [1B1] (ab55178)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [1B1] (ab55178)

      FOXA1 antibody (ab55178) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human salivary gland.

      This image was generated using the ascites version of the product.

    • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [1B1] (ab55178)
      Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [1B1] (ab55178)

      ab55178 at 10 ug/ml staining FOXA1 in human HepG2 cells by Immunocytochemistry/ Immunofluorescence.

      This image was generated using the ascites version of the product.

    • Western blot - Anti-FOXA1 antibody [1B1] (ab55178)
      Western blot - Anti-FOXA1 antibody [1B1] (ab55178)

      FOXA1 antibody (ab55178) at 1ug/lane + HepG2 cell lysate at 25ug/lane.

      This image was generated using the ascites version of the product.

    • Flow Cytometry - Anti-FOXA1 antibody [1B1] (ab55178)
      Flow Cytometry - Anti-FOXA1 antibody [1B1] (ab55178)

      Overlay histogram showing HepG2 cells stained with ab55178 (red line). The cells were fixed with 4 % paraformaldehyde (10 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. The cells were then incubated with the antibody (ab55178, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
      This image was generated using the ascites version of the product.

    文献 (14)

    发表研究结果有使用 ab55178？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab55178 被引用在 14 文献中.

    • Shah SD  et al. Syngeneic model of carcinogen-induced tumor mimics basal/squamous, stromal-rich, and neuroendocrine molecular and immunological features of muscle-invasive bladder cancer. Front Oncol 13:1120329 (2023). PubMed: 36816919
    • Zhang J  et al. Long noncoding RNA LINC01287 promotes proliferation and inhibits apoptosis of lung adenocarcinoma cells via the miR-3529-5p/RNASEH2A axis under the competitive endogenous RNA pattern. Environ Toxicol 36:2093-2104 (2021). PubMed: 34254728
    • Geusz RJ  et al. Sequence logic at enhancers governs a dual mechanism of endodermal organ fate induction by FOXA pioneer factors. Nat Commun 12:6636 (2021). PubMed: 34789735
    • Thanan R  et al. Opposing Roles of FoxA1 and FoxA3 in Intrahepatic Cholangiocarcinoma Progression. Int J Mol Sci 21:N/A (2020). PubMed: 32151057
    • Li H  et al. Olfactomedin 4 mediation of prostate stem/progenitor-like cell proliferation and differentiation via MYC. Sci Rep 10:21924 (2020). PubMed: 33318499
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-4 of 4 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FOXA1 antibody

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (hES derived definitive endoderm)
    Specification
    hES derived definitive endoderm
    Fixative
    Methanol
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Oct 22 2012

    Question

    Dear Sir,   Thanks for a positive reply and for that you are willing to offer a free replacement antibody. After the bad experience with the presently available batch of  ab23728 I would rather avoid using that anymore. Instead I would like to have ab55178 hoping that this will offer a working approach to our experiments in studying FoxA1.  Please do not ship this antibody today but rather wait until Monday April 23 since it may otherwise be held up over the weekend at some airport.   Thank you for taking my complains seriously

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of this product with AB55178. This has been set to ship on Monday with the order number XXXX. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Question

     I was running my experiments with the same reagents comparing the two different FoxA1 antibodies from one day to the next and hence there is no need to troubleshoot. Furthermore,  the same protocol and buffers were also used several fold with other antibodies since then without any problems. Hence the low titer in the second delivery from Abcam is just a very low titer. If you do not want to acknowledge that then there is just a waste of time to dwell more on this issue. If you had really wanted to compare the titers then you should have shown a dilution experiment with the two different lots of antibody and a constant amount of antigen. however, the 100-fold difference that we observed should have been evident in almost any kind of experiment along this line. The data you send me are not informative and raises more questions than answers. The only tissue that seem to be from the same extract is the heart (ab29431) when comparing the two different lots but it remains unclear how much sample that was loaded on the two different gels. Furthermore I do not understand have lanes 3 and 7 can appear so different when the ame antibody was used for probing (ab23738) albeit one was chip grade (lanes 5-8). The ChIP grade antibody does not even recognize any band around 50 KDa (= MW of FoxA1) see lane 7.   One way to  repair these problems, seen from my standpoint, is to give  me a corresponding amount of the old lot (ab 23738, lot 459088) if it still remains in stock or alternatively to send my another FoxA1 antibody (e.g. ab55178).If not then I have to buy a FoxA1 antibody from another company and redirect my future orders accordingly.

    Read More
    Answer

    Thank you for your response.

    As I have stated before, we do take the quality of our products seriously. We no longer have the previous lot in stock. I would be happy to offer a free of charge replacement for this product either of another lot or another product, ab55178. Alternately I may refund the original purchase for you.

    Please let me know how you would like to proceed.

    Read More

    Question

    Dear Sir/Madam,
    I have ordered your Peptide ab23074 (FOXA1 peptide 463-473) but I see
    no signal with your ab55178 (ImmunogenRecombinant fragment,
    corresponding to amino acids 367-473 of Human FOXA1).
    I was very disappointed with this finding, do you have a different
    peptide that would be recognised by this antibody?
    I enclose an attachment with the data showing strong signals with the
    Her2 peptide, but NO signal with the FOXA1 peptide (Cell line used as a
    positive control)
    Kind regards,

    Read More
    Answer

    I am very sorry to hear you are disappointed after using the FOXA1 peptide (aa 463-473, ab23074) with the Anti-FOXA1 antibody (ab55178).

    The antibody is monoclonal antibody that will only bind one epitope in the FOXA1 protein. This antibody was generated with an immunogen corresponding to the amino acids 367-473.

    Since this antibody give a very strong signal with endogenous positive control that was used, Ican explain the lack of signal with the peptide:

    The immunogen is much longer than the peptide. And the antibody therefore binds somewhere between amino acid 376-463.

    Unfortunately, we do not offer the immunogen peptide for ab55178 at the moment in our catalogue.I suggest to use the full length protein (ab98301, )as control if needed. The cell line that was used is also a good control.

    I hope this information is helpful. Please do not hesitate to contact me again with any furtherquestions.

    Read More

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