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Our Abpromise guarantee covers the use of ab4147 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. See Abreview|
|WB||Use a concentration of 0.03 - 0.1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 71 kDa).Can be blocked with Human NUMB peptide (ab22990).|
|IP||1/1000. For 1 hr on mouse brain lysate (see Abreview).|
|ICC/IF||Use at an assay dependent dilution. PubMed: 17127749|
ab4147 staining NUMB in the NIH3T3 cell line from Mouse fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 60 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Donkey anti-rabbit polyclonal(1/500) was used as the secondary antibody.
Western Blot analysis of rat brain lysate (35µg protein in RIPA buffer)labeling NUMB with ab4147 at 0.01µg/ml. Primary incubation was 1 hour. Detected by chemiluminescence.
Single collagenase-isolated mouse muscle fibre stained with DAPI (blue) to reveal nuclei and anti-Numb (green) which detects satellite cells.
This picture was kindly supplied as part of the review submitted by Christopher Mann.
ab4147 staining (0.03µg/ml) of Mouse Brain lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
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