Anti-TRBC1 antibody [EPR24928-52]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TRBC1 antibody [EPR24928-52] (AB325497)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood);Raji (human Burkitt's lymphoma B lymphocyte) cells labelling with ab325497 at 1/1000 (0.507 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 Jurkat cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Negative control : 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-TRBC1 antibody [EPR24928-52] (AB325497)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized HEK-293T(human embryonic kidney epithelial cell) cells transfected with an empty expression vector containing a myc-His-tag®;HEK-293T cells transfected with a human TRBC1 expression vector containing a myc-His-tag®;HEK-293T cells transfected with a human TRBC2 expression vector containing a myc-His-tag®; cells labelling with ab325497 at 1/1000 (0.507 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 293T cells (shown in green) transfected a human TRBC1 expression vectorcontaining a myc-His-tag®, showing no staining in 293T cells transfected a human TRBC2 expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Panel A : HEK-293T cells transfected with an empty expression vector containing a myc-His-tag®; Panel B : HEK-293T cells transfected with a human TRBC1 expression vector containing a myc-His-tag®; Panel C : HEK-293T cells transfected with a human TRBC2 expression vector containing a myc-His-tag®

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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-TRBC1 antibody [EPR24928-52] (AB325497)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized H9(human lymphoma cutaneous T lymphocyte);CCRF-CEM (human peripheral blood T lymphoblast) cells labelling with ab325497 at 1/1000 (0.507 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 and membranous staining in H9 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Negative control : CCRF-CEM(PMID : 29131157). 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-TRBC1 antibody [EPR24928-52] (AB325497)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling with ab325497 at 1/1000 (0.507 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 subsets of CD3+ human PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-human CD3 mouse monoclonal antibody (Alexa Fluor® 647) 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-TRBC1 antibody [EPR24928-52] (AB325497)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling with ab325497 at 1/1000 (0.507 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 subsets of CD20 negative human PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab279298 Anti-CD20 mouse monoclonal antibody - Mouse IgG1 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.

• WB

Lab

Western blot - Anti-TRBC1 antibody [EPR24928-52] (AB325497)

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

Negative control : Ramos, Raji.

CCRF-CEM is a TCR beta-1 negative but TCR beta-2 positive cell line while H9 is a TCR beta-2 negative but TCR beta-1 positive cell line(PMID : 29131157).

The identity of the bands higher than 50 kDa are unknown.

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

All lanes:

Western blot - Anti-TRBC1 antibody [EPR24928-52] (ab325497) at 1/1000 dilution

Lane 1:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 2:

H9 (human lymphoma cutaneous t lymphocyte) whole cell lysate at 20 µg

Lane 3:

CCRF-CEM (human peripheral blood t lymphoblast) whole cell lysate at 20 µg

Lane 4:

HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate at 20 µg

Lane 5:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 6:

Ramos (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 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 1/20000 dilution

Observed band size: 38 kDa,36 kDa

false

Exposure time: 92s

• WB

Lab

Western blot - Anti-TRBC1 antibody [EPR24928-52] (AB325497)

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

Low expression : cerebellum.

The identity of the bands higher than 50 kDa are unknown.

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

All lanes:

Western blot - Anti-TRBC1 antibody [EPR24928-52] (ab325497) at 1/1000 dilution

Lane 1:

Human lymph node tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 38 kDa,36 kDa

false

Exposure time: 114s