Anti-RAB32 antibody [EPR30866-577]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB32 antibody [EPR30866-577] (AB327059)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) Raji (human Burkitt's lymphoma B lymphocyte) cells labelling RAB32 with ab327059 at 1/50 (9.72 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 cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : Raji. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB32 antibody [EPR30866-577] (AB327059)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) HDLM-2 (human Hodgkin lymphoma cell) cells labelling RAB32 with ab327059 at 1/50 (9.72 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 cytoplasmic staining in U-87 MG cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : HDLM-2. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• I-ELISA

Lab

Indirect ELISA - Anti-RAB32 antibody [EPR30866-577] (AB327059)

Indirect ELISA analysis of ab327059 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 RAB32;Human RAB38;Human RAB10;Human RAB29.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Human RAB38, RAB10 and RAB29 by ELISA.

• WB

Lab

Western blot - Anti-RAB32 antibody [EPR30866-577] (AB327059)

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

Low expression : Raji, HDLM-2.

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

All lanes:

Western blot - Anti-RAB32 antibody [EPR30866-577] (ab327059) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

BT-549 (human Breast Mammary gland epithelial) whole cell lysate at 50 µg

Lane 3:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 50 µg

Lane 4:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-RAB32 antibody [EPR30866-577] (AB327059)

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

Low expression : Jurkat.

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

All lanes:

Western blot - Anti-RAB32 antibody [EPR30866-577] (ab327059) at 1/1000 dilution

Lane 1:

A-172 (human brain glioblastoma cell ) whole cell lysate at 50 µg

Lane 2:

HaCaT (human skin keratinocyte) whole cell lysate at 50 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-RAB32 antibody [EPR30866-577] (AB327059)

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

Low expression : cerebellum.

The identity of the higher MW band at approximately 37 kDa (in lane 3) is unknown.

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

All lanes:

Western blot - Anti-RAB32 antibody [EPR30866-577] (ab327059) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 50 µg

Lane 2:

Human colon tissue lysate at 50 µg

Lane 3:

Human cerebellum tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 26 kDa,36 kDa

false

Exposure time: 81s