HNF 4 alpha (HNF4A) (NM_000457) Human Recombinant Protein

CAT#: TP317863

Recombinant protein of human hepatocyte nuclear factor 4, alpha (HNF4A), transcript variant 2, 20 µg

Product Images

In vitro oligonucleotide binding assay. C-myc-tagged HNF4A protein (OriGene TP317863) was added to a solution of double-stranded oligonucleotides labeled with biotin. Oligonucleotides consisted of DNA sequences of wild-type (WT) or HV SLC4A5 alleles. After incubation for 30 min, streptavidin Alexa Fluor 647 was added to label the oligonucleotides; anti-c-Myc antibody was then added, followed by magnetic protein A/G to capture HNF4A-oligonucleotide complexes. The samples were washed and read on a microplate reader. HNF4A binding is increased in HV relative to WT sequence. Figure cited from PLoS ONE, PMID: 29642240

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  Coomassie blue staining of purified HNF4A protein (Cat# TP317863). The protein was produced from HEK293T cells transfected with HNF4A cDNA clone (Cat# RC217863) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Species: Human
• Expression Host: HEK293T
• Expression cDNA Clone or AA Sequence:
  Protein Sequence

  >RC217863 representing NM_000457
  Red=Cloning site Green=Tags(s)
  
  MRLSKTLVDMDMADYSAALDPAYTTLEFENVQVLTMGNDTSPSEGTNLNAPNSLGVSALCAICGDRATGK
  HYGASSCDGCKGFFRRSVRKNHMYSCRFSRQCVVDKDKRNQCRYCRLKKCFRAGMKKEAVQNERDRISTR
  RSSYEDSSLPSINALLQAEVLSRQITSPVSGINGDIRAKKIASIADVCESMKEQLLVLVEWAKYIPAFCE
  LPLDDQVALLRAHAGEHLLLGATKRSMVFKDVLLLGNDYIVPRHCPELAEMSRVSIRILDELVLPFQELQ
  IDDNEYAYLKAIIFFDPDAKGLSDPGKIKRLRSQVQVSLEDYINDRQYDSRGRFGELLLLLPTLQSITWQ
  MIEQIQFIKLFGMAKIDNLLQEMLLGGSPSDAPHAHHPLHPHLMQEHMGTNVIVANTMPTHLSNGQMCEW
  PRPRGQAATPETPQPSPPGGSGSEPYKLLPGAVATIVKPLSAIPQPTITKQEVI
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Tag: C-Myc/DDK
• Predicted MW: 52.6 kDa
• Concentration: >0.05 µg/µL as determined by microplate BCA method
• Purity: > 80% as determined by SDS-PAGE and Coomassie blue staining
• Buffer: 25 mM Tris-HCl, 100 mM glycine, pH 7.3, 10% glycerol
• Bioactivity: HNF4A Activity Verified in a DNA-binding Assay: HNF4A (TP317863, transcript variant 2) activity was measured in a colorimetric DNA-binding assay. Purified HNF4A protein containing a C-terminal MYC/DDK tag was incubated with biotinylated double-stranded oligonucleotide containing the HNF4A consensus DNA-binding sequence (see below). Following incubation, the reaction was transferred to a streptavidin-coated microplate to allow capture of the DNA-protein complex. After washing, the captured protein was detected with an anti-DDK peroxidase conjugate and colorimetric signal detection with TMB. Specificity of the protein-DNA interaction was confirmed by carrying out the binding in the presence of an unlabeled competitor oligonucleotide and by comparison to binding to an oligonucleotide containing a mutation in the consensus binding sequence.
  EMSA assay (PMID: 25598084)
  Binding assay (PMID: 29642240)
• Preparation: Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
• Note: For testing in cell culture applications, please filter before use. Note that you may experience some loss of protein during the filtration process.
• Storage: Store at -80°C.
• Stability: Stable for 12 months from the date of receipt of the product under proper storage and handling conditions. Avoid repeated freeze-thaw cycles.

Reference Data

• RefSeq: NP_000448
• Locus ID: 3172
• UniProt ID: P41235, F1D8T1
• Cytogenetics: 20q13.12
• Refseq Size: 4737
• Refseq ORF: 1422
• Synonyms: FRTS4; HNF4; HNF4a7; HNF4a8; HNF4a9; HNF4alpha; MODY; MODY1; NR2A1; NR2A21; TCF; TCF-14; TCF14
• Summary: The protein encoded by this gene is a nuclear transcription factor which binds DNA as a homodimer. The encoded protein controls the expression of several genes, including hepatocyte nuclear factor 1 alpha, a transcription factor which regulates the expression of several hepatic genes. This gene may play a role in development of the liver, kidney, and intestines. Mutations in this gene have been associated with monogenic autosomal dominant non-insulin-dependent diabetes mellitus type I. Alternative splicing of this gene results in multiple transcript variants encoding several different isoforms. [provided by RefSeq, Apr 2012]
• Protein Families: Druggable Genome, ES Cell Differentiation/IPS, Nuclear Hormone Receptor, Transcription Factors
• Protein Pathways: Maturity onset diabetes of the young