Products:Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> ECM Enzymes >> Kallikreins
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In the datasheet one can find the applicaton note, that it works in western blot. Does this mean under native or under denaturing conditions? Also the Abreview from Douglas Taylor about western blot with this ab doesn´t tell, if it was made under native or denatured conditions. I´m asking, because we are searching now since quite a space of time for an ab against KLK5, which functions in paraffin-embedded tissue sections. Thanks a lot for your assistance and patience. |
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ANSWER: |
Thank you for your enquiry. The antibody ab7283 has been tested in standard reducing conditions in western blotting and has not been tested in IHC-P. It may work in IHC-P and should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will offer you 50 Abpoints which can be redeemed on a number of rewards (a further 100 Abpoints will be offered for an image).
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
IHC image of ab7283 staining Kallikrein 5 in Human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7283, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab7283 stained HeLa cells. The cells were 4% formaldehyde (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 (ab7283, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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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