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Mitofilin Overexpression Lysate

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Western Blot: Mitofilin Overexpression Lysate (Adult Normal) [NBL1-11979] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Mitofilin.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Mitofilin Overexpression Lysate Summary

Description

Mitofilin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC201854

Accession#: NM_006839

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
IMMT

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
83.5 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 Mitofilin Overexpression Lysate

  • DKFZp779P1653
  • inner membrane protein, mitochondrial
  • MGC111146
  • Mic 60
  • Mic60
  • MICOS Complex Subunit MIC60
  • mitochondrial (mitofilin)
  • mitofilin
  • motor protein
  • p87/89
  • proliferation-inducing gene 4

Background

Mitochondria are the center of cellular energy production and essential metabolic reactions. As double membrane-bound organelles, mitochondria from different species, tissues, and metabolic states are highly polymorphic in nature, yet exhibit common structural features. The ultrastructural variations in mitochondrial architecture occur mainly due to the differences in the amount and shape of cristae. Abundant cristae are found in mitochondria from tissues where energy demand is high. Analysis of the human heart mitochondrial proteome shows that mitofilin is one of the most abundant mitochondrial proteins. It appears to play an important role in the maintenance of cristae morphology. Mitofilin was originally described as heart muscle protein (HMP) because of its high expression in the heart.

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.

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Blogs on Mitofilin.

Mitofilin and the Mitochondrial Inner Membrane Organizing System (MINOS)
Mitofilin was originally described as a heart muscle protein due to its high expression in the heart. Recently, analysis of the human heart mitochondrial proteome demonstrated that Mitofilin is one of the most abundant mitochondrial proteins (1). Rese...  Read full blog post.

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Bioinformatics

Gene Symbol IMMT