Overview

  • Product name
  • Description
    Rabbit polyclonal to HEXIM1
  • Host species
    Rabbit
  • Tested applications
    Suitable 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.

    Read Abcam's proprietary immunogen policy (Peptide available as ab26978.)

  • Positive control
    • Recombinant Human HEXIM1 protein (ab117163) can be used as a positive control in WB. This antibody gave 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 (Data not shown): Kidney,Testis , Ovary ; Rat Tissue Lysate (Data not shown): Kidney 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

  • Function
    Transcriptional 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 specificity
    Ubiquitously expressed with higher expression in placenta. HEXIM1 and HEXIM2 are differentially expressed. Expressed in endocrine tissues.
  • Sequence similarities
    Belongs to the HEXIM family.
  • Domain
    The coiled-coil domain mediates oligomerization.
  • Cellular localization
    Nucleus. 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

Images

  • All lanes : Anti-HEXIM1 antibody (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
    All lanes : 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?



    ab25388 recognizes a band at approximately 54 kDa that corresponds in size to that seen for HEXIM1. Although it has a predicted molecular weight of 41 kDa, it has been shown to migrate at a larger size of about 54-60 kDa (see Byers et al., J Biol Chem. 2005 Apr 22;280(16):16360-7 and Schulte et al., J. Biol. Chem., Vol. 280, (26): 24968-24977).
  • 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

This product has been referenced in:
  • Faust TB  et al. The HIV-1 Tat protein recruits a ubiquitin ligase to reorganize the 7SK snRNP for transcriptional activation. Elife 7:N/A (2018). Read more (PubMed: 29845934) »
  • Egloff S  et al. The 7SK snRNP associates with the little elongation complex to promote snRNA gene expression. EMBO J 36:934-948 (2017). Read more (PubMed: 28254838) »
See all 20 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
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)
Negative control
Rabbit control IgG
Specification
breast epithelial MCF7
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control
In-house HEXIM1 antibody

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%

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"

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