Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human thymus tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling MTH1 with ab200832 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of lung tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human squamous cell carcinoma of lung tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Immunocytochemistry/Immunofluorescence analysis of Daudi (human Burkitt's lymphoma) labelling MTH-1 with purified ab200832 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

• IP

Supplier Data

Immunoprecipitation - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

MTH1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200832 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab200832 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : Jurkat whole cell lysate 10ug (Input).
Lane 2 : ab200832 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab200832 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

All lanes:

Immunoprecipitation - Anti-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>)

Predicted band size: 23 kDa

Observed band size: 18 kDa

false

• WB

Supplier Data

Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Blocking/dilution Buffer : 5% NFDM/TBST.

The expression profile observed is consistent with the following literature PMID : 11296483.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

All lanes:

Western blot - Anti-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>) at 1/5000 dilution

Lane 1:

Human fetal kidney lysate at 20 µg

Lane 2:

Human fetal thymus lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 23 kDa

Observed band size: 18 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Blocking/dilution Buffer : 5% NFDM/TBST.

The expression profile observed is consistent with the following literature PMID : 11296483.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

All lanes:

Western blot - Anti-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>) at 1/5000 dilution

All lanes:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 23 kDa

Observed band size: 18 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : MTH1 knockout HAP1 cell lysate (20 μg)
Lane 3 : Jurkat cell lysate (20 μg)
Lane 4 : A549 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab200832 was shown to specifically react with MTH1 when MTH1 knockout samples were used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. ab200832 at a dilution of 1/5000 and ab8245 (loading control to GAPDH) diluted to 1/10,000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

All lanes:

Western blot - Anti-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>)

Predicted band size: 23 kDa

false

• WB

Lab

Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (AB246327)

This data was developed using the same antibody clone in a different buffer formulation (ab200832).

Lanes 1- 4 : Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab200832 was shown to react with MTH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266400 (knockout cell lysate ab257565) was used. Wild-type HEK-293T and NUDT1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab200832 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

NUDT1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nudt1-mth1-knockout-hek-293t-cell-line-ab266400'>ab266400</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 18 kDa

false