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    hexim1-antibody-ab25388.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Anti-HEXIM1 antibody (ab25388)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (23)

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Western blot - Anti-HEXIM1 antibody (ab25388)
  • Immunoprecipitation - Anti-HEXIM1 antibody (ab25388)
  • Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody (ab25388)

Key features and details

  • Rabbit polyclonal to HEXIM1
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-HEXIM1 antibody
    See all HEXIM1 primary antibodies
  • Description

    Rabbit polyclonal to HEXIM1
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • 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.)

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Co-factors
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors

Associated products

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IP
Human
WB
Human
All applications
Mouse
Rat
Dog
Application Abreviews Notes
WB (1)
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.
IP
Use at an assay dependent concentration.
Notes
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.
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.
  • Target information above from: UniProt accession O94992 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10614 Human
    • Entrez Gene: 192231 Mouse
    • Entrez Gene: 498008 Rat
    • Omim: 607328 Human
    • SwissProt: O94992 Human
    • SwissProt: Q8R409 Mouse
    • SwissProt: Q5M9G1 Rat
    • Unigene: 15299 Human
    • Unigene: 586945 Human
    • Unigene: 634944 Human
    • Unigene: 741758 Human
    • Unigene: 8763 Mouse
    • Unigene: 113703 Rat
    see all
  • 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

  • Western blot - Anti-HEXIM1 antibody (ab25388)
    Western blot - Anti-HEXIM1 antibody (ab25388)
    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).
  • Immunoprecipitation - Anti-HEXIM1 antibody (ab25388)
    Immunoprecipitation - Anti-HEXIM1 antibody (ab25388)
    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 .
  • Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody (ab25388)
    Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody (ab25388)

    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.

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (23)

    Publishing research using ab25388? Please let us know so that we can cite the reference in this datasheet.

    ab25388 has been referenced in 23 publications.

    • Edwards DS  et al. BRD4 Prevents R-Loop Formation and Transcription-Replication Conflicts by Ensuring Efficient Transcription Elongation. Cell Rep 32:108166 (2020). PubMed: 32966794
    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
    • Bowry A  et al. BET Inhibition Induces HEXIM1- and RAD51-Dependent Conflicts between Transcription and Replication. Cell Rep 25:2061-2069.e4 (2018). PubMed: 30463005
    • 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). 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). PubMed: 28254838
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    ChIP abreview for Anti-HEXIM1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Dr. Monica Montano

    Verified customer

    Submitted Apr 10 2007

    Western blot abreview for Anti-HEXIM1 antibody

    Excellent
    Abreviews
    Abreviews
    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%
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 05 2007

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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