Anti-TCR beta antibody [EPR24928-66]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCR beta antibody [EPR24928-66] (AB315324)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TRBC1 expression vector containing a Myc-His tag. (B) HEK-293T transfected with a TRBC2 expression vector containing a Myc-His tag. (C) HEK-293T transfected with empty vector containing a Myc-His tag. Tissue labeling TCR beta with ab315324 at 1/5000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TRBC1 expression vector containing a Myc-His tag and (B) HEK-293T transfected with a TRBC2 expression vector containing a Myc-His tag, negative staining on (C) HEK-293T transfected with empty vector containing a Myc-His tag. The section was incubated with ab315324 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCR beta antibody [EPR24928-66] (AB315324)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling TCR beta with ab315324 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver. The section was incubated with ab315324 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

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Multiplex immunohistochemistry - Anti-TCR beta antibody [EPR24928-66] (AB315324)

Fluorescence multiplex immunohistochemical analysis of human tonsil (formalin-fixed paraffin-embedded section). Merged staining of Panel A : merged staining of anti-TCR beta (green; Opal™520) and anti-CD3E (red; Opal™570) on human tonsil, Panel B : anti-TCR beta staining T lymphocytes of human tonsil, Panel C : anti-CD3E staining T lymphocytes of human tonsil. DAPI was used as a counter stain. Followed by Opal Polymer HRP Ms + Rb. 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. The section was incubated in three rounds of staining : in the order of ab315324, ab237707 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval was performed 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 [EPR24928-66] (AB315324)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling TCR beta with ab315324 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen. The section was incubated with ab315324 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCR beta antibody [EPR24928-66] (AB315324)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling TCR beta with ab315324 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung cancer. The section was incubated with ab315324 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCR beta antibody [EPR24928-66] (AB315324)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling TCR beta with ab315324 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The section was incubated with ab315324 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.

• WB

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Western blot - Anti-TCR beta antibody [EPR24928-66] (AB315324)

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

Negative control : Ramos, Raji.

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).

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

All lanes:

Western blot - Anti-TCR beta antibody [EPR24928-66] (ab315324) 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

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: 180s

• WB

Supplier Data

Western blot - Anti-TCR beta antibody [EPR24928-66] (AB315324)

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

Low expression : liver, cerebellum.

The molecular weight observed is consistent with what has been described in the literature (PMID : 31461748, 37796108).

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

All lanes:

Western blot - Anti-TCR beta antibody [EPR24928-66] (ab315324) at 1/1000 dilution

Lane 1:

Human lymph node tissue lysate at 20 µg

Lane 2:

Human thymus tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human cerebellum tissue 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: 36 kDa,38 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TCR beta antibody [EPR24928-66] (AB315324)

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

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.

All lanes:

Western blot - Anti-TCR beta antibody [EPR24928-66] (ab315324) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 10 µg

Lane 2:

293T transfected with a human TCR beta-1 expression vector containing a His-tag, whole cell lysate at 10 µg

Lane 3:

293T transfected with a human TCR beta-2 expression vector containing a His-tag, whole cell lysate at 10 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: 22 kDa,36 kDa

false

Exposure time: 180s