MTHFR Recombinant antibody

MTHFR Uni-rAb^TM Recombinant Antibody for WB, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240348C12

Cat no : 83372-7-RR

Synonyms

Methylenetetrahydrofolate reductase (NADPH), EC:1.5.1.53, 240348C12

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Validation Data Gallery

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  WB analysis using 83372-7-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83372-7-RR (MTHFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  FC experiment of HeLa using 83372-7-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug MTHFR Recombinant antibody (83372-7-RR, Clone:240348C12) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83372-7-RR

  Biolayer interferometry (BLl) kinetic assays of 83372-7-RR against Human MTHFR were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: Daudi cells, Raji cells, Jurkat cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 83372-7-RR is MTHFR Fusion Protein expressed in E. coli.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: MTHFR fusion protein Ag14970
• Full Name: 5,10-methylenetetrahydrofolate reductase (NADPH)
• Calculated Molecular Weight: 656 aa, 75 kDa
• Observed Molecular Weight: 75-78 kDa
• GenBank Accession Number: BC053509
• Gene Symbol: MTHFR
• Gene ID (NCBI): 4524
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

5,10-Methylenetetrahydrofolate reductase (MTHFR) is a crucial enzyme in the folate metabolic pathway with a key role in generating methyl groups. MTHFR reduces 5,10-methylenetetrahydrofolate (5,10-methyleneTHF) to 5-methylTHF, the primary carbon donor for methionine production from homocysteine. In humans and mice studies, mutations and polymorphisms in MTHFR are associated with impaired spermatogenesis and male infertility. It has been reported that MTHFR is highly phosphorylated under unperturbed conditions and T34 is the priming phosphorylation site. MTHFR gene has two promoters and isoform (70 kDa and 77 kDa). Although the major form of MTHFR is 77kDa in most tissues, a second isoform of 70kDa was observed after western blotting of human fetal liver and porcine liver. (PMID: 24769206, PMID: 34128976, PMID: 18615588)