Rabbit Recombinant Monoclonal MTHFR antibody. Carrier free. Suitable for ICC/IF, IP, WB, IHC-P and reacts with Mouse, Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ICC/IF | IP | WB | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Note that mouse was not tested in IP but is predicted to react. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a cosubstrate for homocysteine remethylation to methionine (PubMed:29891918). Represents a key regulatory connection between the folate and methionine cycles (Probable).
Methylenetetrahydrofolate reductase (NADPH), MTHFR
Rabbit Recombinant Monoclonal MTHFR antibody. Carrier free. Suitable for ICC/IF, IP, WB, IHC-P and reacts with Mouse, Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ab251399 is the carrier-free version of Anti-MTHFR antibody [EPR19781] ab203786.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
The 'MTHFR' protein also known as methylenetetrahydrofolate reductase plays an important role in folate metabolism by catalyzing the reduction of 510-methylenetetrahydrofolate to 5-methyltetrahydrofolate. This enzyme has a mass of approximately 70 kDa and is expressed in various tissues including liver kidney and brain. It exists primarily in the cytoplasm where it performs its enzymatic functions critical for cellular metabolism.
Methylenetetrahydrofolate reductase is essential in the process of transferring one-carbon units especially in the synthesis of the amino acid methionine and the methylation of DNA. This enzyme does not form part of a larger protein complex but operates independently in the folate cycle. Its activity directly influences homocysteine levels within the cell with alterations in its function affecting many metabolic processes.
Methylenetetrahydrofolate reductase is an important component of the folate and methionine cycles significant in the remethylation of homocysteine to methionine. These pathways intersect with transmethylation and nucleotide synthesis essential for cell functions and replication. It interacts with proteins like methionine synthase and cystathionine beta-synthase which further demonstrate the enzyme's importance in maintaining cellular homeostasis.
Methylenetetrahydrofolate reductase deficiency or mutations often lead to hyperhomocysteinemia and have been associated with cardiovascular diseases and neurological disorders. Mutations in this enzyme can alter its efficiency leading to the accumulation of homocysteine. This has connections with proteins such as methionine synthase where impaired functioning due to MTHFR mutation can exacerbate the disruption of homocysteine metabolism contributing to disease states.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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Terms & Conditions.
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1: 3 minutes; Lanes 2-4: 30 seconds.
All lanes: Western blot - Anti-MTHFR antibody [EPR19781] (Anti-MTHFR antibody [EPR19781] ab203786) at 1/1000 dilution
Lane 1: Human fetal brain lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Lane 4: Human spleen lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 35 kDa, 75 kDa
Observed band size: 75 kDa
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MTHFR with Anti-MTHFR antibody [EPR19781] ab203786 at 1/1000 (0.509 µg/mL) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on human kidney. The section was incubated with Anti-MTHFR antibody [EPR19781] ab203786 for 30 minutes 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 minutes.
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling MTHFR with Anti-MTHFR antibody [EPR19781] ab203786 at 1/1000 (0.509 µg/mL) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on human thyroid carcinoma. The section was incubated with Anti-MTHFR antibody [EPR19781] ab203786 for 30 minutes 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 minutes.
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Lanes 1 - 3: Western blot - Anti-MTHFR antibody [EPR19781] (Anti-MTHFR antibody [EPR19781] ab203786) at 1/1000 dilution
Lanes 4 - 5: Western blot - Anti-MTHFR antibody [EPR19781] (Anti-MTHFR antibody [EPR19781] ab203786) at 1/5000 dilution
Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2: HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 3: Human fetal liver tissue lysate at 20 µg
Lane 4: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 5: Mouse placenta tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 75 kDa
Observed band size: 75 kDa
Exposure time: 3min
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-MTHFR antibody [EPR19781] (Anti-MTHFR antibody [EPR19781] ab203786) at 1/1000 dilution
Lane 1: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg
Lane 3: Mouse kidney tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 42 kDa, 45 kDa, 75 kDa
Observed band size: 42 kDa, 75 kDa, 90-120 kDa
Exposure time: 3min
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation.MTHFR was immunoprecipitated from 0.35mg of HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate with Anti-MTHFR antibody [EPR19781] ab203786 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-MTHFR antibody [EPR19781] ab203786 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution. Lane 1: HT-29 whole cell lysate, 10µg (Input). Lane 2: Anti-MTHFR antibody [EPR19781] ab203786 IP in HT-29 whole cell lysate. Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-MTHFR antibody [EPR19781] ab203786 in HT-29 whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 10 seconds.
All lanes: Immunoprecipitation - Anti-MTHFR antibody [EPR19781] (Anti-MTHFR antibody [EPR19781] ab203786)
Predicted band size: 75 kDa
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling MTHFR with Anti-MTHFR antibody [EPR19781] ab203786 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: Anti-MTHFR antibody [EPR19781] ab203786 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution. -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
This data was developed using Anti-MTHFR antibody [EPR19781] ab203786, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling MTHFR with Anti-MTHFR antibody [EPR19781] ab203786 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on RAW 264.7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: Anti-MTHFR antibody [EPR19781] ab203786 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution. -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
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