Overview

  • Product name

    HNF1 Transcription Factor Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Nuclear Extracts
  • Assay type

    Semi-quantitative
  • Sensitivity

    300 ng/well
  • Assay time

    3h 30m
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    HNF1 Transcription Factor Assay Kit (Colorimetric) (ab207209) is a high throughput assay to quantify HNF1 activation in nucleat extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.


    A specific double stranded DNA sequence containing the HNF1 consensus binding site (5’ –GTTAATNATTAAC– 3’) has been immobilized onto a 96-well plate. Active HNF1 present in the nuclear extract specifically binds to the oligonucleotide. HNF1 is detected by a primary antibody that recognizes an epitope of HNF1 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout at OD 450 nm. This product detects mouse, rat and human HNF1.


    Key performance and benefits:



    • Assay time: 3.5 hours (cell extracts preparation not included).

    • Detection limit: < 0.3 µg nuclear extract/well.

    • Detection range: 0.3 – 5 µg nuclear extract/well.

  • Notes

    HNF1 (Hepatocyte Nuclear Factor-1) is a protein involved in liver development, metabolism and glucose homeostasis. HNF1α and 1β (vHNF1) are two related transcription factors of the homeodomain family. Unlike other family members, HNF1α and 1β have an extra 21 amino acid DNA binding segment in their homeobox, and a POU A-related domain that is involved in DNA binding. The N-terminus contains a dimerization domain allowing for homo- or heterodimerization. There are also three C-terminus activation domains that are rich in proline and glycine, proline and glutamine, and serine, respectively. HNF1α is located on chromosome 12 and HNF1β is found on chromosome 17.

    HNF1 was originally discovered in the liver, where it was thought to be restricted. Later, it was also found to play important roles in the kidney, small intestine, thymus and pancreas. The HNF1 consensus-binding site is the inverted palindrome GTTAATNATTAAC. HNF1 can bind to this sequence as a homodimer, but other factors, such as NF-1 or C/EBP, are required for efficient transcriptional activity. HNF1 plays a crucial role in liver development during embryogenesis and metabolism homeostasis in the adult. It helps regulate such genes as glucose-cotransporter-2 in the kidney, phenylalanine hydroxylase (PAH) in the liver, and insulin, L-pyruvate kinase and aldolase B in pancreatic B cells.

    Mutations in HNF1 have been implicated in the early onset of type II diabetes referred to as MODY (maturity onset diabetes of the young). MODY is an inherited disease characterized by early onset of diabetes mellitus. MODY3 and 5 represent mutations in genes encoding HNF1α and 1β, respectively. The most common form is MODY3, which accounts for about 60% of cases, while MODY5 is relatively rare. In all forms of MODY, insulin levels are affected by defective secretions of pancreatic B-cells.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests 5 x 96 tests
    10X Antibody Binding Buffer 1 x 2.2ml 1 x 11ml
    10X Wash Buffer 1 x 22ml 1 x 110ml
    96-well HNF-1 assay plate 1 unit 5 units
    Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 1 x 11µl 1 x 55µl
    Binding Buffer 1 x 10ml 1 x 50ml
    Developing Solution 1 x 11ml 1 x 55ml
    Dithiothreitol (DTT) (1 M) 1 x 100µl 1 x 500µl
    Hep G2 nuclear extract (2.5 μg/μL) 1 x 40µl 1 x 200µl
    HNF 1 antibodies 1 x 11µl 1 x 55µl
    Lysis Buffer 1 x 10ml 1 x 50ml
    Mutated oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl
    Plate sealer 1 unit 5 units
    Protease Inhibitor Cocktail 1 x 100µl 1 x 500µl
    Stop Solution 1 x 11ml 1 x 55ml
    Wild-type oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl
  • Research areas

  • Function

    Transcriptional activator that regulates the tissue specific expression of multiple genes, especially in pancreatic islet cells and in liver. Required for the expression of several liver specific genes. Binds to the inverted palindrome 5'-GTTAATNATTAAC-3'.
  • Tissue specificity

    Liver.
  • Involvement in disease

    Defects in HNF1A are a cause of hepatic adenomas familial (HEPAF) [MIM:142330]. Hepatic adenomas are rare benign liver tumors of presumable epithelial origin that develop in an otherwise normal liver. Hepatic adenomas may be single or multiple. They consist of sheets of well-differentiated hepatocytes that contain fat and glycogen and can produce bile. Bile ducts or portal areas are absent. Kupffer cells, if present, are reduced in number and are non-functional. Conditions associated with adenomas are insulin-dependent diabetes mellitus and glycogen storage diseases (types 1 and 3). Note=Bi-allelic inactivation of HNF1A, whether sporadic or associated with MODY3, may be an early step in the developmant of some hepatocellular carcinomas.
    Defects in HNF1A are the cause of maturity-onset diabetes of the young type 3 (MODY3) [MIM:600496]; also symbolized MODY-3. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
    Defects in HNF1A are the cause of susceptibility to diabetes mellitus insulin-dependent type 20 (IDDM20) [MIM:612520]. IDDM20 is a multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These features can result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
  • Sequence similarities

    Belongs to the HNF1 homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Alternative names

    • Albumin proximal factor
    • Hepatic nuclear factor 1
    • Hepatic nuclear factor 1 alpha
    • Hepatic transcription factor 1
    • Hepatic transcription factor 1 alpha
    • Hepatocyte nuclear factor 1-alpha
    • HNF 1
    • HNF 1A
    • HNF-1-alpha
    • HNF-1A
    • hnf1a
    • HNF1A_HUMAN
    • Interferon production regulator factor
    • LF B1
    • LF B1 hepatic nuclear factor
    • LFB 1
    • LFB1
    • LFB1 hepatic nuclear factor
    • Liver specific transcription factor LF B1
    • Liver specific transcription factor LFB1
    • Liver-specific transcription factor LF-B1
    • Maturity onset diabetes of the young 3
    • MODY 3
    • MODY3
    • TCF 1
    • TCF-1
    • TCF1
    • Transcription factor 1
    • Transcription factor 1 hepatic
    see all
  • Database links

Images

  • Different amounts of unstimulated HeLa (grey) and HepG2 (black) were tested for HNF1 activation. These results are provided for demonstration purposes only.

Protocols

References

ab207209 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab207209.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up