Question (9226) | Anti-Aurora A antibody (ab1287)

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Question

BATCH NUMBER 62608 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No nuclear staining seen, only cytoplasmic staining. SAMPLE Tissue, PRIMARY ANTIBODY Abcam ab1287, 1/250 in NSS/Tris buffered saline, 40mins, rtemp, wash for 5 mins. SECONDARY ANTIBODY Dako Duet Kit-K0492, as advised in kit DETECTION METHOD Dako Duet Kit-K0492, as advised in kit POSITIVE AND NEGATIVE CONTROLS USED Trying to get staining on breast and colon tumour. ANTIBODY STORAGE CONDITIONS As advised. FIXATION OF SAMPLE Neutral buffered formalin ANTIGEN RETRIEVAL Heat mediated, using pressure cooker and microwave techniques (separately) BLOCKING CONDITIONS Normal swine serum

Answer

Thank you for your enquiry and for sending us your detailed protocol. We would suggest blocking the non-specific binding sites with 1%BSA for 1 hr and incubating the samples with the primary antibody for 2 hrs (40 min is too short). This antibody ahs been tested for IHC using human pancreatic tumor sections, so it may be worth using this type of tissue sections as positive control. Should you still have problem with this antibody, please do not hesitate to contact us again.

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