Anti-HEXIM1 antibody (ab25388)
Key features and details
- Rabbit polyclonal to HEXIM1
- Suitable for: WB, ICC/IF, IP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-HEXIM1 antibody
See all HEXIM1 primary antibodies -
Description
Rabbit polyclonal to HEXIM1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIP HumanWB Human
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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
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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
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
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
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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
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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
see all -
Alternative names
- Cardiac lineage protein 1 antibody
- CLP 1 antibody
- CLP1 antibody
see all
Images
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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)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)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
References (23)
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
