Recombinant Anti-CHD4 antibody [EPR22953-38] - ChIP Grade (ab240640)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22953-38] to CHD4 - ChIP Grade
- Suitable for: IP, Flow Cyt (Intra), WB, ICC/IF, ChIP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-CHD4 antibody [EPR22953-38] - ChIP Grade
See all CHD4 primary antibodies -
Description
Rabbit monoclonal [EPR22953-38] to CHD4 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: IP, Flow Cyt (Intra), WB, ICC/IF, ChIPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NCCIT, K562, HeLa, HEK-293 and RAW264.7 lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells. IP: NCCIT cells. ChIP: Chromatin prepared from K562 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22953-38 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab240640 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 |
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IP |
1/30.
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Flow Cyt (Intra) |
1/600.
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WB |
1/1000. Predicted molecular weight: 218 kDa.
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ICC/IF |
1/500.
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ChIP |
Use 5 µg for 25 µg of chromatin.
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Notes |
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IP
1/30. |
Flow Cyt (Intra)
1/600. |
WB
1/1000. Predicted molecular weight: 218 kDa. |
ICC/IF
1/500. |
ChIP
Use 5 µg for 25 µg of chromatin. |
Target
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Function
Component of the histone deacetylase NuRD complex which participates in the remodeling of chromatin by deacetylating histones. -
Sequence similarities
Belongs to the SNF2/RAD54 helicase family.
Contains 2 chromo domains.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 2 PHD-type zinc fingers. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. Cytoplasm > cytoskeleton > centrosome. Associates with centrosomes in interphase. - Information by UniProt
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Database links
- Entrez Gene: 1108 Human
- Entrez Gene: 107932 Mouse
- Omim: 603277 Human
- SwissProt: Q14839 Human
- SwissProt: Q6PDQ2 Mouse
- Unigene: 162233 Human
- Unigene: 333388 Mouse
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Alternative names
- ATP dependent helicase CHD4 antibody
- ATP-dependent helicase CHD4 antibody
- CHD 4 antibody
see all
Images
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All lanes : Anti-CHD4 antibody [EPR22953-38] - ChIP Grade (ab240640) at 1/1000 dilution
Lane 1 : NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate
Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 5 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 218 kDa
Observed band size: 280 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: PMID: 28572115) .
Exposure time: 1 second.
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Chromatin was prepared from K-562 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab240640 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are from paper PMID: 23505388.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
CHD4 was immunoprecipitated from 0.35 mg NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10ug with ab240640 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240640 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: NCCIT whole cell lysate 10ug.
Lane 2: ab240640 IP in NCCIT whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240640 in NCCIT whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CHD4 with ab240640 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling CHD4 with ab240640 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear staining and weak cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling CHD4 with ab240640 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear staining and weak cytoplasmic staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab240640 has not yet been referenced specifically in any publications.