Overview

  • Product name
    Anti-MTHFR antibody [EPR19781]
    See all MTHFR primary antibodies
  • Description
    Rabbit monoclonal [EPR19781] to MTHFR
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Human MTHFR aa 1-300. The exact sequence is proprietary.
    Database link: P42898

  • Positive control
    • WB: Human fetal brain, fetal heart, fetal kidney, spleen and fetal liver lysates; Jurkat, HT-29, HEK-293, RAW 264.7 and NIH/3T3 whole cell lysates; Mouse placenta and kidney lysates. ICC/IF: Jurkat and RAW 264.7 cells. IP: HT-29 whole cell lysate.
  • General notes

     

     

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab203786 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP 1/30.

Note that mouse was not tested in IP but is predicted to react.

ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 75 kDa).

Target

  • Function
    Catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a co-substrate for homocysteine remethylation to methionine.
  • Pathway
    One-carbon metabolism; tetrahydrofolate interconversion.
  • Involvement in disease
    Defects in MTHFR are the cause of methylenetetrahydrofolate reductase deficiency (MTHFRD) [MIM:236250]. MTHFRD is autosomal recessive disorder with a wide range of features including homocysteinuria, homocysteinemia [MIM:603174], developmental delay, severe mental retardation, perinatal death, psychiatric disturbances, and later-onset neurodegenerative disorders.
    Defects in MTHFR may be a cause of susceptibility to ischemic stroke (ISCHSTR) [MIM:601367]; also known as cerebrovascular accident or cerebral infarction. A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors.
    Defects in MTHFR may be a cause of susceptibility to folate-sensitive neural tube defects (folate-sensitive NTD) [MIM:601634]. The most common NTDs are open spina bifida (myelomeningocele) and anencephaly.
  • Sequence similarities
    Belongs to the methylenetetrahydrofolate reductase family.
  • Information by UniProt
  • Database links
  • Alternative names
    • 5 10 methylenetetrahydrofolate reductase (NADPH) antibody
    • 5 10 methylenetetrahydrofolate reductase antibody
    • Methylenetetrahydrofolate reductase (NAD(P)H) antibody
    • Methylenetetrahydrofolate reductase antibody
    • Methylenetetrahydrofolate reductase intermediate form antibody
    • MTHFR antibody
    • MTHR_HUMAN antibody
    see all

Images

  • All lanes : Anti-MTHFR antibody [EPR19781] (ab203786) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2,3 and 4: 30 seconds.

  • Lanes 1-3 : Anti-MTHFR antibody [EPR19781] (ab203786) at 1/1000 dilution
    Lanes 4-5 : Anti-MTHFR antibody [EPR19781] (ab203786) at 1/5000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 3 : Human fetal liver lysate
    Lane 4 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 5 : Mouse placenta lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : 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
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 3 : Mouse kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 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 ab203786 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (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 (ab7291) at 1/1000 dilution and 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: ab203786 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • 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 ab203786 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (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 (ab7291) at 1/1000 dilution and 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: ab203786 at 1/100 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • MTHFR was immunoprecipitated from 0.35mg of HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate with ab203786 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab203786 at 1/500 dilution.

    VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

    Lane 1: HT-29 whole cell lysate, 10µg (Input).

    Lane 2: ab203786 IP in HT-29 whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab203786 in HT-29 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

This product has been referenced in:
  • Wu JL  et al. MTHFR c.677C>T Inhibits Cell Proliferation and Decreases Prostate Cancer Susceptibility in the Han Chinese Population in Shanghai. Sci Rep 6:36290 (2016). WB ; Human . Read more (PubMed: 27819322) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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