Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

Order now and get it on Tuesday March 02, 2021

Anti-MTH1 antibody [EPR15934-50] (ab200832)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR15934-50] to MTH1
Suitable for: WB, IHC-P, IP, Flow Cyt, ICC/IF
Knockout validated
Reacts with: Human

Product name

Anti-MTH1 antibody [EPR15934-50]
See all MTH1 primary antibodies
Description

Rabbit monoclonal [EPR15934-50] to MTH1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T, HAP1, HeLa and Jurkat whole cell lysate; Human fetal kidney and fetal thymus lysates. IHC-P: Human thymus and squamous cell carcinoma of lung tissues. ICC/IF: Jurkat and A549 cells. Flow Cyt: Jurkat cells. IP: Jurkat whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR15934-50
Isotype

IgG
Research areas

Western blot - Anti-MTH1 antibody [EPR15934-50] (ab200832)

All lanes : Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : NUDT1 knockout HEK-293T cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 23 kDa
Observed band size: 18 kDa
why is the actual band size different from the predicted?

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.
Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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
Western blot - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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.
Western blot - Anti-MTH1 antibody [EPR15934-50] (ab200832)

Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution + Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary
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 why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/dilution Buffer: 5% NFDM/TBST.

The expression profile observed is consistent with the following literature PMID: 11296483.
Western blot - Anti-MTH1 antibody [EPR15934-50] (ab200832)

All lanes : Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal thymus lysate

Lysates/proteins at 20 µg per lane.

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 why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/dilution Buffer: 5% NFDM/TBST.

The expression profile observed is consistent with the following literature PMID: 11296483.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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.
Immunoprecipitation - Anti-MTH1 antibody [EPR15934-50] (ab200832)

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.
Anti-MTH1 antibody [EPR15934-50] (ab200832)

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