Anti-TCPTP antibody [EPR28199-34]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PTPN2 KO Jurkat (human PTPN2 knockout T cell leukemia T lymphocyte from peripheral blood, Left) / Parental Jurkat (Right) cells labelling TCPTP with ab314496 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling TCPTP with ab314496 at 1/500 (1.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly nuclear staining on human colon (PMID : 33001862). The section was incubated with ab314496 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-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded (A) Parental Jurkat (human T cell leukemia) cell pellet (B) PTPN2 knockout Jurkat cell pellet tissue labeling TCPTP with ab314496 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on (A) Parental Jurkat (human T cell leukemia) cell pellet, negative staining on (B) PTPN2 knockout Jurkat cell pellet. The section was incubated with ab314496 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-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling TCPTP with ab314496 at 1/500 (1.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly nuclear staining on rat cardiac muscle. The section was incubated with ab314496 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-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling TCPTP with ab314496 at 1/500 (1.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly nuclear staining on mouse colon. The section was incubated with ab314496 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-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling TCPTP with ab314496 at 1/500 (1.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly nuclear staining on mouse cardiac muscle. The section was incubated with ab314496 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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling TCPTP with ab314496 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling TCPTP with ab314496 at 1/500 (1.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly nuclear staining on rat colon. The section was incubated with ab314496 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

Lab

Western blot - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Western blot : Anti-PTPN2 antibody [EPR28199-34] (ab314496) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab314496 was shown to bind specifically to PTPN2. A band was observed at 48 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PTPN2 knockout cell line. To generate this image, wild-type and PTPN2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TCPTP antibody [EPR28199-34] (ab314496) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN2 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ptpn2-knockout-hct116-cell-line-ab287729'>ab287729</a>)

Lane 2:

PTPN2 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type Jurkat cell lysate at 20 µg

Lane 4:

PTPN2 knockout Jurkat cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1 : 15 seconds Lane 2 : 26 seconds Lane 3 : 92 seconds

All lanes:

Western blot - Anti-TCPTP antibody [EPR28199-34] (ab314496) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

NR8383 (rat lung macrophage (alveolar)) 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 1/100000 dilution

Observed band size: 45 kDa,48 kDa

false

• WB

Supplier Data

Western blot - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression molecular weight observed is consistent with what has been described in the literature (PMID : 25311528 ). Exposure time : Lane 1, 2 : 26 seconds Lane 3 : 59 seconds Lane 4 : 114 seconds

All lanes:

Western blot - Anti-TCPTP antibody [EPR28199-34] (ab314496) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

MEF (mouse embryo fibroblast) 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 1/100000 dilution

Observed band size: 45 kDa,48 kDa

false

• WB

Supplier Data

Western blot - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression molecular weight observed is consistent with what has been described in the literature (PMID : 25311528 ). In Western blot, ab314496 was shown to bind specifically to PTPN2 . Target of interest was observed at 48, 45kDa in wild-type Jurkat cell lysates (lane 1) with no signal observed at this size in PTPN2 knockout cell line (lane 2) (lane 2, knockout cell line ab274899/ knockout cell lysate ab274957). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lane 1, 2 : 3 minutes Lane 3 : 48 seconds

All lanes:

Western blot - Anti-TCPTP antibody [EPR28199-34] (ab314496) at 1/1000 dilution

Lane 1:

Wild-type Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 2:

PTPN2 Knockout Jurkat whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), 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 1/100000 dilution

Observed band size: 45 kDa,48 kDa

false

• WB

Supplier Data

Western blot - Anti-TCPTP antibody [EPR28199-34] (AB314496)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression molecular weight observed is consistent with what has been described in the literature (PMID : 25311528 ).

All lanes:

Western blot - Anti-TCPTP antibody [EPR28199-34] (ab314496) at 1/1000 dilution

Lane 1:

Rat heart tissue lysate at 20 µg

Lane 2:

Rat kidney tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse heart 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 1/100000 dilution

Observed band size: 45 kDa,48 kDa

false

Exposure time: 48s