Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human liver tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression tissue : no staining on human liver. The section was incubated with ab313743 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling METTL16 with ab313743 at 1/50 dilution (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). 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon carcinoma. The section was incubated with ab313743 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human testis tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human testis. The section was incubated with ab313743 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling METTL16 with ab313743 at 1/50 (10.62 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 nuclear staining in HeLa cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse pancreatic tumor tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse pancreatic tumor. The section was incubated with ab313743 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab313743 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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling METTL16 with ab313743 at 1/50 (10.62 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 nuclear staining in Raw264.7 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling METTL16 with ab313743 at 1/100 (5.31 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab313743 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

Supplier Data

Western blot - Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-METTL16 antibody [EPR27236-78] (<a href='/en-us/products/primary-antibodies/mettl16-antibody-epr27236-78-ab313743'>ab313743</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Hela transfected with siRNA specifically targeti METTL16 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: 75 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : liver (PMID : 33930289). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 92 seconds

All lanes:

Western blot - Anti-METTL16 antibody [EPR27236-78] (<a href='/en-us/products/primary-antibodies/mettl16-antibody-epr27236-78-ab313743'>ab313743</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human liver 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: 75 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : liver (PMID : 33930289). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-METTL16 antibody [EPR27236-78] (<a href='/en-us/products/primary-antibodies/mettl16-antibody-epr27236-78-ab313743'>ab313743</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat testis 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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-METTL16 antibody [EPR27236-78] - BSA and Azide free (AB313744)

This data was developed using ab313743, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : liver (PMID : 33930289). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-METTL16 antibody [EPR27236-78] (<a href='/en-us/products/primary-antibodies/mettl16-antibody-epr27236-78-ab313743'>ab313743</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) 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: 75 kDa

false

Exposure time: 10s