Product Review: NUP62 antibody – N-terminal region (ARP46179_P050) in HeLa cells using Immunohistochemistry

Product Page Link: NUP62 antibody – N-terminal region (ARP46179_P050)

Data provided by Dr. Michael Matunis of John Hopkins University

Protocol:

1. Wash HeLa cells 1X with PBS

2. Add fix/perm solution (20 mM PIPES pH=6.8, 4% Formaldehyde, 0.2% TNX100, 10 mM EGTA, 1 mM MgCl2) and let incubate for 10 min. at RT

3. Wash 2X with PBS

4. Put in primary antibody diluted into 2% BSA + 0.02% Sodium Azide in PBS for 1 hour (Nup35 at 2ug/ml; Nup50 at 0.25 ug/ml; Nup62 at 0.25 ug/m; Nup98 at 2ug/ml)

5. Wash 9X (3X in 3 different beakers) of PBS + 0.05% Tween20 

6. Put in secondary Antibody for 30 min. (Invitrogen AlexaFluor488 1:300)

7. Wash 9X (3X in 3 different beakers) of PBS + 0.05% Tween20 

8. Add a mounting solution with DAPI and seal slides.

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