Cytochrome P450 3A5 Overexpression Lysate

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Western Blot: Cytochrome P450 3A5 Overexpression Lysate (Adult Normal) [NBL1-09693] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Cytochrome P450 3A5.

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Summary
Reactivity HuSpecies Glossary
Applications WB
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Cytochrome P450 3A5 Overexpression Lysate Summary

Description

Cytochrome P450 3A5 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC207432

Accession#: NM_000777

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
CYP3A5

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
56.9 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 Cytochrome P450 3A5 Overexpression Lysate

  • aryl hydrocarbon hydroxylase
  • CP35
  • CYP3A5
  • CYPIIIA5
  • Cytochrome P450 3A5
  • Cytochrome P450 HLp2
  • cytochrome P-450
  • cytochrome P450, family 3, subfamily A, polypeptide 5
  • cytochrome P450, subfamily IIIA (niphedipine oxidase), polypeptide 5
  • Cytochrome P450-PCN3
  • DKFZp686L16231
  • EC 1.14.14.1
  • flavoprotein-linked monooxygenase
  • microsomal monooxygenase
  • niphedipine oxidase
  • P450PCN3FLJ31317
  • PCN3
  • xenobiotic monooxygenase

Background

This gene,CYP3A5, encodes a member of the cytochrome P450 superfamily of enzymes. The cytochrome P450 proteins are monooxygenases which catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids and other lipids. This protein localizes to the endoplasmic reticulum and its expression is induced by glucocorticoids and some pharmacological agents. The enzyme metabolizes drugs such as nifedipine and cyclosporine as well as the steroid hormones testosterone, progesterone and androstenedione. This gene is part of a cluster of cytochrome P450 genes on chromosome 7q21.1. This cluster includes a pseudogene, CYP3A5P1, which is very similar to CYP3A5. This similarity has caused some difficulty in determining whether cloned sequences represent the gene or the pseudogene. [provided by RefSeq]. 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.

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Bioinformatics

Gene Symbol CYP3A5