Muscle Phosphofructokinase/PFKM/PFK-1 Overexpression Lysate

Western Blot: PFK-1 Overexpression Lysate (Adult Normal) [NBL1-14316] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PFK-1.

• Reactivity: Hu
• Applications: WB

Muscle Phosphofructokinase/PFKM/PFK-1 Overexpression Lysate Summary

• Description:

  Muscle Phosphofructokinase/PFKM/PFK-1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200702
  
  Accession#: NM_000289
  
  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: PFKM

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: 85 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 Muscle Phosphofructokinase/PFKM/PFK-1 Overexpression Lysate

• 6-phosphofructokinase, muscle type
• EC 2.7.1,6-phosphofructo-1-kinase
• EC 2.7.1.11
• GSD7
• MGC8699
• Muscle Phosphofructokinase
• PFK1
• PFK-1
• PFKA
• PFK-A
• PFKM
• PFKX
• Phosphofructo-1-kinase isozyme A
• Phosphofructokinase 1
• phosphofructokinase, muscle
• phosphofructokinase, polypeptide X
• Phosphofructokinase-1
• Phosphofructokinase-M
• Phosphohexokinase

Background

The PFKM gene encodes the muscle isoform of phosphofructokinase (PFK) (ATP:D-fructose-6-phosphate-1-phosphotransferase, EC 2.7.1.11). PFK catalyzes the irreversible conversion of fructose-6-phosphate to fructose-1,6-bisphosphate and is a key regulatory enzyme in glycolysis. Mammalian PFK is a tetramer made up of various combinations of 3 subunits: muscle (PFKM), liver (PFKL; MIM 171860), and platelet (PFKP; MIM 171840), the genes for which are located on chromosomes 12q13, 21q22, and 10p, respectively. The composition of the tetramers differs according to the tissue type. Muscle and liver PFK are a homotetramers of 4M and 4L subunits, respectively. Erythrocytes contain both L and M subunits, which randomly tetramerize to form M4, L4, and M3L, M2L2, and ML3 hybrid forms of the holoenzyme (Vora et al., 1980 [PubMed 6444721]; Raben and Sherman, 1995 [PubMed 7550225]).[supplied by OMIM]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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: PFKM