Anti-PAX3 抗体 (ab15717)

Thank you for your enquiry and your interest in our products. This product (ab15717) is expected to recognize all 7 isoforms based on the immunizing peptide. However, in WB we only see a 30-35 kDa band in human kidney lysates. I can confirm that ab15717 works in IHC-P as well (tested on formalin-fixed paraffin embedded human oesophagus tissue). In order to be able to distinguish between the different isoforms, Iso-Electric Focusing (IEF) technique can be applied. If you need any further assistance in the future, please do not hesitate to contact me.

Thank you for contacting us. The protocol used to test this antibody in IHC-P is as follows: *Buffers* Tris/ EDTA buffer at pH 9.0:
-Tris 1.21 g
-EDTA 0.37 g
-Distilled water 1000 ml (100 ml to make 10x, 50 ml to make 20x)
Mix to dissolve. pH is usually at 9.0. Add 0.5 ml of Tween 20 and mix well.
Store at room temperature for 3 months or at 4°C for longer storage. *1.1 Deparaffinise and Rehydrate Tissue Sections. * - Remove wax and bring the sections to water. *1.2 **Microwave Antigen Retrieval.* - Place slides in a microwaveable pot containing Tris/ EDTA buffer at pH 9.0, ensure slides are completely covered with solution. - Put pot and slides in microwaveable pressure cooker and place in microwave. - When the cooker has been heated up to pressure, “cook” for 1 - 3 minutes (as appropriate). - Remove the pressure cooker and allow to cool. - When the pressure has dropped, remove the pot containing the slides, and allow the slides to cool/rest. - Wash for a minimum of 5 minutes in tap water. - Wash for 5 minutes in distilled water. *1.3 **Blocking Endogenous Peroxidase.* - Treat sections with a H202 solution ( e.g. 1ml H202 20% in 250ml dH20) and incubate for 15. - Wash slides in PBS *1.4 **Primary Antibody Incubation.* - Apply primary antibody diluted to a concentration of 3ug/mL using 10% NHS in PBS. - Incubate slides at room temperature in a humidity staining chamber for 30 minutes. - Wash in PBS for 2 x 5 minutes. *1.5 **Secondary Antibody Incubation.* - Apply secondary antibody (anti-goat HRP) and incubate slides at room temperature in a humidity staining chamber for 30 minutes. - Wash in PBS for 2 x 5 minutes. *1.6 **Detection.* - Add DAB solution for 5 minutes. - Replace PBS with distilled water to wash for at least 15 minutes (change water frequently). - Counterstain with haematoxylin. Wash in tap water for at least 15 minutes and mount slides with aquamount I hope this helps, please let me know if you have any additional questions.