MTHFD2 Overexpression Lysate

Western Blot: MTHFD2 Overexpression Lysate (Adult Normal) [NBL1-13364] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MTHFD2.

• Reactivity: Hu
• Applications: WB

MTHFD2 Overexpression Lysate Summary

• Description:

  MTHFD2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC203095
  
  Accession#: NM_001040409
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: MTHFD2

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 26.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for MTHFD2 Overexpression Lysate

• bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase
• methenyltetrahydrofolate cyclohydrolase
• methylene tetrahydrofolate dehydrogenase (NAD+ dependent)
• methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2
• mitochondrial
• NAD-dependent methylene tetrahydrofolate dehydrogenase cyclohydrolase
• NMDMC

Background

This gene encodes a nuclear-encoded mitochondrial bifunctional enzyme with methylenetetrahydrofolate dehydrogenase and methenyltetrahydrofolate cyclohydrolase activities. The enzyme functions as a homodimer and is unique in its absolute requirement for magnesium and inorganic phosphate. Formation of the enzyme-magnesium complex allows binding of NAD. Alternative splicing results in multiple transcripts encoding different isoforms. This gene has a pseudogene on chromosome 7. [provided by RefSeq]. Transcript Variant: This variant (2) contains an alternate exon in the 5' coding region and uses a downstream start codon, compared to variant 1. The encoded protein (isoform B) has a shorter N-terminus which lacks a mitochondrial transit peptide and a tetrahydrofolate dehydrogenase/cyclohydrolase catalytic domain, compared to isoform A.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: MTHFD2