Anti-SHMT2/SHMT antibody [EPR28607-51]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney (PMID : 33066813). The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on human cardiac muscle. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on human skeletal muscle. The section was incubated with ab316328 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)

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Flow Cytometry (Intracellular) - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SHMT2/SHMT with ab316328 at 1/500 dilution (0.1 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SHMT2/SHMT with ab316328 at 1/1000 (0.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mitochondrial staining in HeLa cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Red). 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 (2 ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon carcinoma. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hepatocellular carcinoma. The section was incubated with ab316328 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

• IP

Supplier Data

Immunoprecipitation - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

SHMT2/SHMT was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab316328 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316328 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab316328 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316328 in HeLa whole cell lysate

All lanes:

Immunoprecipitation - Anti-SHMT2/SHMT antibody [EPR28607-51] (ab316328) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat liver. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat kidney. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney. The section was incubated with ab316328 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling SHMT2/SHMT with ab316328 at 1/1000 (0.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mitochondrial staining in Hepa1-6 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Red). 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 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on rat skeletal muscle. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling SHMT2/SHMT with ab316328 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on mouse skeletal muscle. The section was incubated with ab316328 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-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Western blot : Anti-SHMT2/SHMT antibody [EPR28607-51] ab316328 staining at 1/5000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 55 kDa in Wild-type A549 cell lysates with no signal observed at this size in SHMT2 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (ab316328) at 1/5000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human SHMT2 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-shmt2-knockout-a549-cell-line-ab301223'>ab301223</a>) at 20 µg

Lane 3:

HeLa at 20 µg

Secondary

Lanes 1 - 3:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 3:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 56 kDa

Observed band size: 55 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

All lanes:

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (ab316328) at 1/5000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Human liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 3:

Rat liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 4:

Rat kidney tissue lysate at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

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

Observed band size: 55 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

Low expression : skeletal muscle

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (ab316328) at 1/5000 dilution

Lane 1:

Hela (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting SHMT2 whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

Mouse kidney tissue lysate at 20 µg with 5% NFDM/TBST

Lane 5:

Mouse liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 6:

Mouse skeletal muscle tissue lysate at 20 µg with 5% NFDM/TBST

Lane 7:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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: 55 kDa,124 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (AB316328)

This antibody does not cross-react with human SHMT1.

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

All lanes:

Western blot - Anti-SHMT2/SHMT antibody [EPR28607-51] (ab316328) at 1/5000 dilution

Lane 1:

His-tagged human SHMT2 recombinant protein at 10 ng with 5% NFDM/TBST

Lane 2:

His-tagged human SHMT1 recombinant protein at 10 ng with 5% NFDM/TBST

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Observed band size: 55 kDa

false

Exposure time: 1s