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One of our customers wants to order ab28252 or ab28253. The tested applications of them are IHC, IP, WB etc in your on-line datasheets. I want to know if they could be applied to IHC-Paraffin.
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ANSWER: |
Thank you for your enquiry. Yes, the antibodies can be used in IHC-P and I have updated the datasheet accordingly. Please use them at an assay dependent dilution using heat-mediated antigen retrieval in citrate buffer pH 6.0. I would recommend a starting dilution of 1:100 although this dilution may have to be optimized. I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
ICC/IF image of ab28252 stained HeLa 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 (ab28252, 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.
All lanes : Anti-Calpastatin antibody - Aminoterminal end (Domain L) Testis Isoform (ab28252) at 1/5000 dilution
Lane 1 : Mouse skeletal tissue muscle lysate
Lane 2 : Mouse skeletal tissue muscle lysate
Lane 3 : Mouse skeletal tissue muscle lysate
Lysates/proteins at 30 µg per lane.
Secondary
HRP-conjugated Goat IgG to Rabbit
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 76 kDa
Observed band size : 25,35,74 kDa (why is the actual band size different from the predicted?)
Lanes 1-3 are skeletal muscle samples from different mice.
This image is courtesy of an anonymous Abreview.
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