Anti-RAB43 antibody [EPR30492-530]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB43 antibody [EPR30492-530] (AB325751)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SR (human pleural effusion lymphoblast) and HeLa (human cervical adenocarcinoma epithelial cell) cells labelling RAB43 with ab325751 at 1/50 (10.34 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing golgi staining in SR cell line (shown in green), which is co-stained with a golgi marker. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Low expression : HeLa.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain golgi at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325751 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab169276 at 1/200 dilution, followed by ab150081 at 1/1000 dilution.

• WB

Lab

Western blot - Anti-RAB43 antibody [EPR30492-530] (AB325751)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HeLa, Daudi.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RAB43 antibody [EPR30492-530] (ab325751) at 1/1000 dilution

Lane 1:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg

Lane 2:

SR (human pleural effusion lymphoblast) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 5:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 23 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-RAB43 antibody [EPR30492-530] (AB325751)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RAB43 antibody [EPR30492-530] (ab325751) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting RAB43 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 23 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-RAB43 antibody [EPR30492-530] (AB325751)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lane 5 of this blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RAB43 antibody [EPR30492-530] (ab325751) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 48 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 48 µg

Lane 3:

Mouse testis tissue lysate at 48 µg

Lane 4:

Mouse stomach tissue lysate at 48 µg

Lane 5:

Mouse brain tissue lysate at 48 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 23 kDa,36 kDa

true

Exposure time: 180s

• I-ELISA

Lab

Indirect ELISA - Anti-RAB43 antibody [EPR30492-530] (AB325751)

Indirect ELISA analysis of ab325751 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human RAB43 protein;Human RAB30 protein;Human RAB19 protein;Human RAB11A protein;Human RAB11B protein.

Antigen concentration : 1000 ng/ml