Overview

  • Product name
  • Description
    Goat polyclonal to HNF-1B
  • Host species
    Goat
  • Specificity
    This antibody is expected to recognize both reported isoforms
  • Tested applications
    Suitable for: WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Dog, Pig, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human HNF-1B aa 248-261 (Cysteine residue).
    Sequence:

    QAYDRQKNPSKEER

  • Positive control
    • Human Liver lysate
  • General notes

    Previously labelled as HNF1 beta. 

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris buffered saline, pH 7.3
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab59118 in the following tested applications.

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

Application Abreviews Notes
WB Use a concentration of 0.1 - 0.3 µg/ml. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
ELISA 1/64000.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Transcription factor, probably binds to the inverted palindrome 5'-GTTAATNATTAAC-3'.
  • Involvement in disease
    Defects in HNF1B are the cause of renal cysts and diabetes syndrome (RCAD) [MIM:137920]; also called maturity-onset diabetes of the young type 5 (MODY5) or familial hypoplastic glomerulocystic kidney disease (GCKD). RCAD is an autosomal dominant disorder comprising non-diabetic renal disease resulting from abnormal renal development, and diabetes, which in some cases occurs earlier than age 25 years and is thus consistent with a diagnosis of maturity-onset diabetes of the young (MODY5). The renal disease is highly variable and includes renal cysts, glomerular tufts, aberrant nephrogenesis, primitive tubules, irregular collecting systems, oligomeganephronia, enlarged renal pelves, abnormal calyces, small kidney, single kidney, horseshoe kidney, and hyperuricemic nephropathy.
    Defects in HNF1B may be rare genetic risk factor contributing to the development of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853]. NIDDM is characterized by an autosomal dominant mode of inheritance, onset during adulthood and insulin resistance.
    Defects in HNF1B may be a cause of susceptibility to prostate cancer hereditary type 11 (HPC11) [MIM:611955]. It is a condition associated with familial predisposition to cancer of the prostate. Most prostate cancers are adenocarcinomas that develop in the acini of the prostatic ducts. Other rare histopathologic types of prostate cancer that occur in approximately 5% of patients include small cell carcinoma, mucinous carcinoma, prostatic ductal carcinoma, transitional cell carcinoma, squamous cell carcinoma, basal cell carcinoma, adenoid cystic carcinoma (basaloid), signet-ring cell carcinoma and neuroendocrine carcinoma.
  • Sequence similarities
    Belongs to the HNF1 homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • FJHN antibody
    • Hepatocyte nuclear factor 1 beta antibody
    • Hepatocyte nuclear factor 1-beta antibody
    • HNF 1B antibody
    • HNF 2 antibody
    • HNF-1-beta antibody
    • HNF-1B antibody
    • HNF1 beta antibody
    • HNF1 homeobox B antibody
    • HNF1B antibody
    • HNF1B_HUMAN antibody
    • HNF1beta antibody
    • HNF2 antibody
    • Homeoprotein LF B3 antibody
    • Homeoprotein LFB3 antibody
    • HPC11 antibody
    • LF B3 antibody
    • LFB3 antibody
    • MODY 5 antibody
    • MODY5 antibody
    • TCF 2 antibody
    • TCF 2 protein antibody
    • TCF-2 antibody
    • TCF2 antibody
    • TCF2 protein antibody
    • Transcription factor 2 antibody
    • Transcription factor 2 hepatic antibody
    • Variant hepatic nuclear factor 1 antibody
    • Variant hepatic nuclear factor antibody
    • VHNF 1 antibody
    • vHNF1 antibody
    see all

Images

  • Anti-HNF-1B antibody (ab59118) at 0.1 µg/ml + Human liver lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 61 kDa
    Observed band size: 61 kDa
    Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.



    The 100kDa band observed was successfully blocked by incubation with the immunizing peptide, however its identity is not known.
  • Definitive endoderm derived from culturing human embryonic stem cells (HUES4) with Inhibin beta a (ab50051) and Wnt3a (ab81484) was further differentiated to primitive gut tube as described in Kroon et al, Nat Biotechnol. (2008). KGF (ab9601) was added to the media to direct differentiation towards primitive gut tube. These images were taken 3 days after addition of KGF, where the primitive gut tube marker TCF2 (ab59118 in green) is present. Dapi staining is shown in blue. Magnification of top panels: 100x, bottom panels: 200x. Negative controls of fluorescently conjugated secondary antibodies only, showed no staining. For more information on this immunofluorescence, please see abreview.

    See Abreview

References

ab59118 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for contacting us.

We are happy to help customers find the most suitable products for their research purposes; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. I have done a precursory search and found two products guaranteed in ICC/IF which may suit your needs. These products are:ab41898 Anti-HNF4 antibody [K9218] (https://www.abcam.com/HNF4-antibody-K9218-ChIP-Grade-ab41898.html) and ab59118Anti-TCF2 antibody (https://www.abcam.com/TCF2-antibody-ab59118.html). While these may be good alternatives we encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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Question

DESCRIPTION OF THE PROBLEM Non-specific staining (TCF2 is a nuclear protein.But we can’t get any anti-TCF2 signal in nuclear.Our result shows this anti-TCF2 antibody has very strong non-spicific binding signal in cytosol of human hepatocyte- HepG2.) SAMPLE Species : Human Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: Cells in culture Sample preparation: HepG2 cells were fixed with 3.7% paraformadehyde PRIMARY ANTIBODY Concentration or dilution 1:200 Diluent buffer PBS Incubation time overnight Buffer: PBST Number of washes: 3 DETECTION METHOD Confocal scaning POSITIVE AND NEGATIVE CONTROLS USED Positive control: Human liver cells-HepG2 ANTIBODY STORAGE CONDITIONS -20°C FIXATION OF SAMPLE If yes: Fixative agent and concentration: 3.7% paraformadehyde Fixation time: 5 min Fixation temperature: room temperature ANTIGEN RETRIEVAL No PERMEABILIZATION STEP Permeabilization method: 5% BAS with 0.5% Triton-X100 at RT 30 min Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Permeabilizing agent and concentration: 0.5% Triton-X100 in 5% BSA BLOCKING CONDITIONS Concentration 5% Blocking time 1 hour Blocking temperature RT SECONDARY ANTIBODY Species: donkey Reacts against: goat Concentration or dilution 1:500 Diluent buffer PBS Incubation time : 1 hour Fluorochrome or enzyme conjugate Alesa-488 Buffer: PBST Number of washes: 3 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Change the secondary antibody dilution from 1:500 to 1: 1000

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Answer

Thank you for sending your customer's protocol and data. The protocol is similar to the protocol used in a positive review on our datasheet from February 2011, using different cells (human embryonic stem cells). I do not have suggestions to improve the protocol but if your customer is confident that HepG2 express this protein, TCF2, and they want to try this antibody again, I I suggest they try fixing the cells with a different fixative, for instance cold methanol for 10 minutes. However, this is just a suggestion; we do not have any data that indicates this will imrove the result. If your customer would like to try a different lot of the antibody, I can send a vial free of charge. We do not have another TCF2 antibody that has been tested for ICC/IF, so a different lot of ab59118 is the only option we have available, other than a credit.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (primitive gut tube from human embryonic stem cells)
Permeabilization
Yes - 0.1% triton
Specification
primitive gut tube from human embryonic stem cells
Blocking step
Dako Protein Block (0.25% caesin) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C
Fixative
Paraformaldehyde

Ms. Emily Moorefield

Verified customer

Submitted Feb 24 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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