Customer Reviews for RAB35 Antibody (ARP52268_P050) tested with HeLa and MEF cells in Western blot

CAT#ARP52268_P050

RAB35 antibody

RAB35 antibody Western Blot

RAB35 antibody HeLa and MEF cells

How do Aviva’s reagents play a role in your experimental goals?

As to Rab35; Detect human and/or mouse Rab35 proteins with WB and IF

How many different experimental trials were conducted using the antibody sample?

once

What type of experimental sample are you using and how did you prepare it?

HeLa and MEF cells. Lysis with lysis buffer (50 mM Tris, 150 mM NaCl, 1% NP40, 2 mM EDTA, 1 mM PMSF, protease inhibitors), took supernatant of 15000rpm centrifugation.
Add 6x SDS‐PAGE sample buffer and boil at 70C for 10 min

What applications did you test the antibody in? Please include dilutions of the primary and secondary reagents.

For Western blot ;
1st Ab, 1:500 dilution in PBS (no milk, no detergent – should be very mild condition). 2nd Ab, anti‐rabbit HRP, 1:10000

What controls were used in your experiment? Please include your positive control.

HeLa cells, HEK293T cells (derived from human)
MEF cells (derived from mouse)
No transfection/GFP‐humanRab35 overexpression / GFP‐mouseSlc3a1 or Slc3a2 overexpression
The PVDF membrane after the experiment with Aviva’s antibody was stripped, and then reprobed with anti‐GFP antibody from another
company.

How did you store the antibody after re‐suspension?

4C

How would you rate this antibody on a scale from 1‐5? Why?

ARP52268:4 Suitable for WB (However, this antibody was not recognized Rab35 by IF.)

Would you use this antibody in future experiments?

Yes

Have you used another antibody which has worked in your application?

Yes. We know another antibody (anti‐humanRab35) can detect human and mouse Rer1 proteins with WB, but it could not be detected endogeneous Rab35 in this test. We are looking for a anti‐Rab35 antibody that can use for IF.

Do you believe the information about the reagent on Aviva’s website is correct?

Yes

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?

Yes.

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