Anti-Rab3A antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab3A antibody (AB3335)

Immunofluorescence analysis of RAB3A was done on 70% confluent log phase HeLa (Human epithelial adenocarcinoma cell line)
cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ab3335 at 2 μg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution for 45 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin at 1/300 dilution. Panel d is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

• WB

Lab

Western blot - Anti-Rab3A antibody (AB3335)

Western blot : Left : Anti-RAB3A antibody (ab3335) staining at 2 ug/ml. Right : Recombinant Anti-Rab3A antibody [EPR26022-9] (ab302518) staining at 1/1000 dilution. Anti-Rab3A antibodies shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab3335 and ab302518 were shown to bind specifically to RAB3A. A band was observed at 27 kDa in wild-type SK-N-FI cell lysates with no signal observed at this size in RAB3A knockout cell line. To generate this image, wild-type and RAB3A knockout SK-N-FI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-Rab3A antibody (ab3335) at 2 µg/mL

Lanes 1 - 4:

Western blot - Anti-Rab3A antibody [EPR26022-9] (<a href='/ko/products/primary-antibodies/rab3a-antibody-epr26022-9-ab302518'>ab302518</a>) at 1/1000 dilution

Lane 1:

Wild-type SK-N-FI cell lysate at 20 µg

Lane 2:

RAB3A knockout SK-N-FI cell lysate at 20 µg

Lane 3:

Human brain cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Observed band size: 27 kDa

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