Immunocytochemistry/ Immunofluorescence: Chd7 Antibody [NBP1-77393] - Antibody was tested in NIH/3T3 cells with FITC (green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red).
Immunohistochemistry: Chd7 Antibody [NBP1-77393] - IHC analysis of CHD7 in mouse intestine using DAB with hematoxylin counterstain.
This CHD7 antibody is useful for IHC-P, ICC/IF and Western blot where a band is seen ~330 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Human and mouse. Immunogen has 85% identity to chicken Chd7.
Packaging, Storage & Formulations
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
PBS and 30% Glycerol
0.05% Sodium Azide
Immunogen affinity purified
Manufactured by Genomic Antibody Technology™. GAT FAQs
Alternate Names for CHD7 Antibody
ATP-dependent helicase CHD7
chromodomain helicase DNA binding protein 7 isoform CRA_e
chromodomain helicase DNA binding protein 7
chromodomain-helicase-DNA-binding protein 7
CHARGE syndrome (coloboma, heart defects, atresia of the choanae, retarded growth and development, genital hypoplasia, ear anomalies and deafness) is a congenital malformation syndrome caused by mutations in the CHD7 (chromodomain helicase DNA-binding protein) gene in approximately 2/3 of cases. In Kallmann syndrome, a similar proportion of affected individuals also have mutated CHD7. These mutations probably affect neurogenerative anomalies and maturation events through SOX2 interaction. Expression patterns of CHD7 in combination with SOX2 evaluation can provide some insight into molecular causes of CHARGE and Kallmann syndromes.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Fixation: 4% paraformaldehyde in PBS pH 7.4 for 15 minutes at room temperature Permeabilization: PBS containing 0.2% Triton X-100 for 10 minutes Blocking: PBST (PBS+ 0.1% Tween 20) 10% goat serum for 60 minutes Immunostaining: primary Ab 1:100 in PBST in a humidified chamber overnight at 4°C
Product General Protocols
View specific protocols for CHD7 Antibody (NBP1-77393):
FAQs for CHD7 Antibody (NBP1-77393). (Showing 1 - 1 of 1 FAQs).
I am working with Chd7 and have just visited your website. I would like to have some advices or protocols to use the NBP1-77393 Ab, especially western blot protocol/extraction buffers. My concern is that Chd7 is a high molecular weight protein and it seems very difficult to extract it and to transfer it on a membrane I was just wondering if you had any tips regarding this matter
It was a 5% gel ran 100V for 60 min stressing that it was done in cold conditions with ice packs. It should be ok to do overnight at a lower voltage also in cold conditions. We had the best success on NIH3T3 cells, but also tested on Jurkat. In regards to your inquiry about the Western Blot protocol specific for NBP1-77393, we do have one available on our website. You can find this under the protocol tab. In case you have trouble locating this I will provide it to you in this email. These are just suggestions and some additional optimization may need to be done to achieve the best results based on sample preparation and protein expression. Western Blot Protocol: 1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane. 2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus. 3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. 4. Rinse the blot. 5. Block the membrane using standard blocking buffer for at least 1 hour. 6. Wash the membrane in wash buffer three times for 10 minutes each. 7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature. 8. Wash the membrane in wash buffer three times for 10 minutes each. 9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature. 10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background). 11. Apply the detection reagent of choice in accordance with the manufacturers instructions. Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
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