Overview

  • Product nameAnti-HEXIM1 antibody - ChIP Grade
    See all HEXIM1 primary antibodies
  • Description
    Rabbit polyclonal to HEXIM1 - ChIP Grade
  • Tested applicationsSuitable for: ChIP, WB, ICC/IF, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human HEXIM1.

    (Peptide available as ab26978.)

  • Positive control
    • This antibody gave a positive signal in the following Human Whole Cell Lysates: HeLa Jurkat A431 HEK293 (Data not shown) MCF7 SHSY-5Y Mouse Whole Cell Lysate MEF1 NIH 3T3 (Data not shown) Mouse Tissue Lysates Kidney (Data not shown) Testis (Data not shown) Ovary (Data not shown) Rat Tissue Lysate Kidney (Data not shown) This antibody gave a positive signal in the following Methanol/Formaldehyde fixed cell line: HeLa.

Properties

Applications

Our Abpromise guarantee covers the use of ab25388 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
ChIP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 41 kDa). Abcam recommends using milk (3%) as the blocking agent.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • FunctionTranscriptional regulator which functions as a general RNA polymerase II transcription inhibitor. In cooperation with 7SK snRNA sequesters P-TEFb in a large inactive 7SK snRNP complex preventing RNA polymerase II phosphorylation and subsequent transcriptional elongation. May also regulate NF-kappa-B, ESR1, NR3C1 and CIITA-dependent transcriptional activity.
  • Tissue specificityUbiquitously expressed with higher expression in placenta. HEXIM1 and HEXIM2 are differentially expressed. Expressed in endocrine tissues.
  • Sequence similaritiesBelongs to the HEXIM family.
  • DomainThe coiled-coil domain mediates oligomerization.
  • Cellular localizationNucleus. Cytoplasm. Binds alpha-importin and is mostly nuclear.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cardiac lineage protein 1 antibody
    • CLP 1 antibody
    • CLP1 antibody
    • EDG 1 antibody
    • EDG1 antibody
    • Estrogen down-regulated gene 1 protein antibody
    • FLJ13562 antibody
    • Hexamethylene bis acetamide inducible 1 antibody
    • Hexamethylene bis acetamide inducible protein antibody
    • Hexamethylene bis acetamide inducible transcript 1 antibody
    • Hexamethylene bis-acetamide-inducible protein 1 antibody
    • HEXI1_HUMAN antibody
    • HEXIM 1 antibody
    • Hexim1 antibody
    • HEXIM1 protein antibody
    • HIS 1 antibody
    • HIS1 antibody
    • HMBA inducible antibody
    • MAQ 1 antibody
    • MAQ1 antibody
    • Menage a quatre 1 antibody
    • Menage a quatre protein 1 antibody
    • Protein HEXIM1 antibody
    see all

Anti-HEXIM1 antibody - ChIP Grade images

  • All lanes : Anti-HEXIM1 antibody - ChIP Grade (ab25388) at 1/250 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 5 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 41 kDa
    Observed band size : 54 kDa (why is the actual band size different from the predicted?)
  • HEXIM1 - ChIP Grade was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to HEXIM1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab25388.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 54kD: HEXIM1 .
  • Image courtesy of Human Protein Atlas

    ab25388 staining HEXIM1 in human kidney. Paraffin embedded human kidney tissue was incubated with ab25388 (1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab25388 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • ICC/IF image of ab25388 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab25388 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-HEXIM1 antibody - ChIP Grade (ab25388)

This product has been referenced in:
  • Mück F  et al. DDX6 transfers P-TEFb kinase to the AF4/AF4N (AFF1) super elongation complex. Am J Blood Res 6:28-45 (2016). Read more (PubMed: 27679741) »
  • Liu P  et al. Release of positive transcription elongation factor b (P-TEFb) from 7SK small nuclear ribonucleoprotein (snRNP) activates hexamethylene bisacetamide-inducible protein (HEXIM1) transcription. J Biol Chem 289:9918-25 (2014). WB, ChIP ; Human . Read more (PubMed: 24515107) »

See all 11 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (breast epithelial MCF7)
Specification breast epithelial MCF7
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step Semiquantitative PCR
Positive control In-house HEXIM1 antibody
Negative control Rabbit control IgG
Username

Dr. Monica Montano

Verified customer

Submitted Apr 10 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (cervical cancer cells (HeLa))
Loading amount 20 µg
Specification cervical cancer cells (HeLa)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Username

Abcam user community

Verified customer

Submitted Feb 05 2007

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