Anti-HEXIM1 antibody (ab25388)
- Datasheet
- Specific References (16)
- Protocols
Overview
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Product nameAnti-HEXIM1 antibody
See all HEXIM1 primary antibodies -
DescriptionRabbit polyclonal to HEXIM1
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Host speciesRabbit
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Tested applicationsSuitable for: ChIP, WB, ICC/IF, IHC-P, IPmore details
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Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Dog
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human HEXIM1.
(Peptide available as ab26978.)
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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
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Positive Controls
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
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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.
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Tissue specificityUbiquitously expressed with higher expression in placenta. HEXIM1 and HEXIM2 are differentially expressed. Expressed in endocrine tissues.
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Sequence similaritiesBelongs to the HEXIM family.
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DomainThe coiled-coil domain mediates oligomerization.
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Cellular localizationNucleus. 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 . -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXIM1 antibody (ab25388)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
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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
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) »
- Albert TK et al. The Establishment of a Hyperactive Structure Allows the Tumour Suppressor Protein p53 to Function through P-TEFb during Limited CDK9 Kinase Inhibition. PLoS One 11:e0146648 (2016). Read more (PubMed: 26745862) »
Customer reviews and Q&As
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