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ab4959 |
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Do you have any images of a western using this antibody and the predicted molecular weight? Thank you for your help!
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ANSWER: |
Thank you for your enquiry. We currently do not have an image of the western blot with ab3350, all the information, including images, we have for all our products is listed on the on-line datasheets for your convenience and we update these as soon as we get more data. Ab3350 was tested on hamster BMAL1/aryl hydrocarbon nuclear translocator 3 (ARNT3)fusion construct overexpressed in E. coli (a 110 kDa protein) as well as recombinant human BMAL1. The predicted MW of BMAL1 is 69.4 kDa. Please do not hesitate to contact us again if you need further assistance,
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Anti-BMAL1 antibody - ChIP Grade (ab3350) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Predicted band size : 69.4 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
Additional bands at : 51 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 3 minutes
The 75 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to BMAL1.
ICC/IF image of ab3350 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab3350, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.
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