Overview

  • Product name

    Elk1 Transcription Factor Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Nuclear Extracts
  • Assay type

    Semi-quantitative
  • Sensitivity

    < 1000 ng/well
  • Assay time

    3h 30m
  • Species reactivity

    Reacts with: Human
  • Product overview

    Elk1 Transcription Factor Assay Kit (Colorimetric) (ab207202) is a high throughput assay to quantify Elk1 activation in nuclear 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 Elk1 consensus binding site has been immobilized onto a 96-well plate. Active Elk1 present in the nuclear extract specifically binds to the oligonucleotide. Elk1 is detected by a primary antibody that recognizes an epitope of Elk1 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout that at OD 450 nm. This product detects only human Elk1.


    Key performance and benefits:



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

    • Detection limit: < 1.0 µg nuclear extract/well.

    • Detection range: 1.0 – 10 µg nuclear extract/well.

  • Notes

    Elk1 is a transcription factor that is activated by all three MAPK pathways. It is a member of the ternary complex factor (TCF) subgroup of the Ets protein family, along with Sap1 and NET/ERP/SAP2/Elk-3. Ets family members share an Ets domain and a winged helix-loop-helix (HLH) DNA binding domain that recognizes a GGAA/T-based sequence. TCFs are able to form a ternary complex with the serum response factor (SRF) and the serum-response element (SRE), and are involved in SRE-driven gene expression.

    Elk1 has three major functional domains. The N-terminal Ets-DNA binding domain recognizes GGAA/T sequences. A motif of 20 amino acids, called the B domain, mediates protein-protein interactions with SRF. The C-terminal portion of Elk-1 has phosphorylation sites for ERK, JNK and p38 MAPKs. Elk-1 is mainly activated by ERK proteins, and binds to the SRE present in many immediate early genes such as c-fos, egr1, egr2, pip92 and nur77

  • 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 assay plate 1 unit 5 units
    Anti-mouse 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
    HeLa (TPA, 10 min) nuclear extract (2.5 μg/μL) 1 x 40µl 1 x 200µl
    Lysis Buffer 1 x 10ml 1 x 50ml
    Mutated oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl
    Phosphorylated Elk-1 antibody 1 x 11µl 1 x 55µl
    Plate sealer 1 unit 5 units
    Poly [d(l-c)] (17 µg/μL) 1 x 100µl 1 x 500µl
    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

    Stimulates transcription. Binds to purine-rich DNA sequences. Can form a ternary complex with the serum response factor and the ETS and SRF motifs of the fos serum response element.
  • Tissue specificity

    Lung and testis.
  • Sequence similarities

    Belongs to the ETS family.
    Contains 1 ETS DNA-binding domain.
  • Post-translational
    modifications

    Sumoylation represses transcriptional activator activity as it results in recruitment of HDAC2 to target gene promoters which leads to decreased histone acetylation and reduced transactivator activity. It also regulates nuclear retention.
    On mitogenic stimulation, phosphorylated on C-terminal serine and threonine residues by MAPK1. Ser-383 and Ser-389 are the preferred sites for MAPK1. In vitro, phosphorylation by MAPK1 potentiates ternary complex formation with the serum responses factors, SRE and SRF. Phosphorylation leads to loss of sumoylation and restores transcriptional activator activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Alternative names

    • ELK 1
    • Elk1
    • ELK1 member of ETS oncogene family
    • ELK1 protein
    • ELK1, ETS transcription factor
    • ELK1_HUMAN
    • ELK2 member of ETS oncogene family
    • ETS domain containing protein Elk 1
    • ETS domain containing protein Elk1
    • ETS domain protein Elk1
    • ETS domain-containing protein Elk-1
    • ETS like gene 1
    • Member of ETS oncogene family
    • Oncogene Elk1
    • Tyrosine kinase (ELK1) oncogene
    see all
  • Database links

Images

  • Different amounts of nuclear extracts from untreated U-937 (light grey), untreated HeLa (dark grey) and HeLa treated with TPA for 10 min (black) cells were tested for Elk-1 activity. These results are provided for demonstration purposes only.

Protocols

References

ab207202 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab207202.
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