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If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab22512 for help.
Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
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I am using the Anti-WDR5 antibody (ab22512) and my Western Blots are not looking as good as what you have shown. I was wondering if you could send me the conditions you have used? |
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ANSWER: |
Thank you for your enquiry. I am sorry to hear ab22512 is giving you trouble. I don't have a specific protocol to use with this antibody but if you could let me know some more about your sample preparation and experimental protocol I'd be happy to offer any tips I can. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - WDR5 antibody (ab22512)
All lanes : Anti-WDR5 antibody (ab22512) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole cell lysate
Lane 3 : Brain (Rat) Whole Cell Lysate - normal tissue
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Predicted band size : 36 kDa
Observed band size : 36 kDa
Additional bands at : 30 kDa,48 kDa (possible post-translational modification),55-60 kDa. We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - Anti-WDR5 antibody (ab22512)
ICC/IF image of ab22512 stained Hek293 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 ab22512 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.
Western blot - Anti-WDR5 antibody (ab22512)
All lanes : Anti-WDR5 antibody (ab22512) at 1 µg/ml
Lane 1 : Marker
Lane 2 : Zebrafish brain homogenate at 20 µg
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 36 kDa
Observed band size : 37 kDa (why is the actual band size different from the predicted?)
Exposure time : 5 minutes
Anti-WDR5 antibody for Western blot in Mouse (22512)
WDR5 antibody for Western blot in Fruit fly (Drosophila melanogaster) (22512)
Anti-WDR5 antibody for Western blot in Human (22512)
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