Anti-TCR beta 抗体 [RM2056]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TCR beta antibody [RM2056] (AB318205)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining CD3 with ab21703 at a 1/150 (0.06 ug/ml) dilution, ab318205 anti-TCR beta used at 1/500 (1.036 ug/ml) dilution and ab64088 anti-CD20 used at a 1/50 (0.2 ug/ml) dilution.

Panel A : merged staining of anti-CD3 (magenta; Opal™690), anti-TCR beta (green; Opal™520) and anti-CD20 (yellow; Opal™570) on human tonsil.
Panel B : anti-CD3 staining T lymphocytes in human tonsil.
Panel C : anti-TCR beta staining T lymphocytes in human tonsil.
Panel D : anti-CD20 staining B lymphocytes in human tonsil.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab21703, ab318205 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TCR beta antibody [RM2056] (AB318205)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining CD3 with ab21703 at a 1/150 (0.06 ug/ml) dilution, ab318205 anti-TCR beta used at 1/500 (1.036 ug/ml) dilution and ab64088 anti-CD20 used at a 1/50 (0.2 ug/ml) dilution.

Panel A : merged staining of anti-CD3 (magenta; Opal™690), anti-TCR beta (green; Opal™520) and anti-CD20 (yellow; Opal™570) on human colon.
Panel B : anti-CD3 staining T lymphocytes in human colon.
Panel C : anti-TCR beta staining T lymphocytes in human colon.
Panel D : anti-CD20 staining B lymphocytes in human colon.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab21703, ab318205 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TCR beta antibody [RM2056] (AB318205)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD3 with ab21703 at a 1/150 (0.06 ug/ml) dilution, ab318205 anti-TCR beta used at 1/500 (1.036 ug/ml) dilution and ab64088 anti-CD20 used at a 1/50 (0.2 ug/ml) dilution.

Panel A : merged staining of anti-CD3 (magenta; Opal™690), anti-TCR beta (green; Opal™520) and anti-CD20 (yellow; Opal™570) on human spleen.
Panel B : anti-CD3 staining T lymphocytes in human spleen.
Panel C : anti-TCR beta staining T lymphocytes in human spleen.
Panel D : anti-CD20 staining B lymphocytes in human spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab21703, ab318205 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCR beta antibody [RM2056] (AB318205)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling TCR beta with ab318205 at 1/500 (1.036 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen (PMID : 33867734).
The section was incubated with ab318205 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TCR beta antibody [RM2056] (AB318205)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD3 with ab21703 at a 1/150 (0.06 ug/ml) dilution, ab318205 anti-TCR beta used at 1/500 (1.036 ug/ml) dilution and ab64088 anti-CD20 used at a 1/50 (0.2 ug/ml) dilution.

Panel A : merged staining of anti-CD3 (magenta; Opal™690), anti-TCR beta (green; Opal™520) and anti-CD20 (yellow; Opal™570) on mouse spleen.
Panel B : anti-CD3 staining T lymphocytes in mouse spleen.
Panel C : anti-TCR beta staining T lymphocytes in mouse spleen.
Panel D : anti-CD20 staining B lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab21703, ab318205 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TCR beta antibody [RM2056] (AB318205)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat spleen tissue staining CD3 with ab21703 at a 1/150 (0.06 ug/ml) dilution, ab318205 anti-TCR beta used at 1/500 (1.036 ug/ml) dilution and ab64088 anti-CD20 used at a 1/50 (0.2 ug/ml) dilution.

Panel A : merged staining of anti-CD3 (magenta; Opal™690), anti-TCR beta (green; Opal™520) and anti-CD20 (yellow; Opal™570) on rat spleen.
Panel B : anti-CD3 staining T lymphocytes in rat spleen.
Panel C : anti-TCR beta staining T lymphocytes in rat spleen.
Panel D : anti-CD20 staining B lymphocytes in rat spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab21703, ab318205 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-TCR beta antibody [RM2056] (AB318205)

Negative control : Ramos, Raji, skeletal muscle, cerebellum.

This antibody recognizes both TCR beta-1 and TCR beta-2 .

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 molecular weight observed is consistent with what has been described in the literature (PMID : 31461748).

Lanes 1-5 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 6-8 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-3, 5-8 : 92 seconds; Lane 4 : 15 seconds.

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

All lanes:

Western blot - Anti-TCR beta antibody [RM2056] (ab318205) at 1/1000 dilution

Lane 1:

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

Lane 2:

Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

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

Lane 6:

Human lymph node tissue lysate at 20 µg

Lane 7:

Human skeletal muscle tissue lysate at 20 µg

Lane 8:

Human cerebellungm tissue lysate at 20 µg

Secondary

Lanes 1 - 5:

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

Lanes 6 - 8:

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

Observed band size: 38 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-TCR beta antibody [RM2056] (AB318205)

Negative control : A20, skeletal muscle.

The molecular weight observed is consistent with what has been described in the literature (PMID : 31461748).

The identity of the lower MW band at approximately 30kDa in lane 8 is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-7 : 48 seconds; Lane 8 : 6 seconds.

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

All lanes:

Western blot - Anti-TCR beta antibody [RM2056] (ab318205) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 2:

A20 (mouse reticum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

Mouse lymph node tissue lysate at 20 µg

Lane 4:

Mouse thymus tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Rat spleen tissue lysate at 20 µg

Lane 7:

Rat skeletal muscle tissue lysate at 20 µg

Lane 8:

Rat thymus tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 38 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-TCR beta antibody [RM2056] (AB318205)

This antibody recognizes both TCR beta-1 and TCR beta-2.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

Exposure time : Lanes 1-3 : 48 seconds; Lanes 4-5 : 10 seconds.

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

All lanes:

Western blot - Anti-TCR beta antibody [RM2056] (ab318205) at 1/1000 dilution

Lane 1:

293T transfected with an empty vector containing a his tag whole cell lysate at 20 µg

Lane 2:

293T transfected with a human TCR beta-1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a human TCR beta-2 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T transfected with a mouse TCR beta-1 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 5:

293T transfected with a mouse TCR beta-2 expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 22 kDa,36 kDa

false