Aromatase Overexpression Lysate

Images

 
Western Blot: Aromatase Overexpression Lysate (Adult Normal) [NBP2-11585] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Aromatase.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

Aromatase Overexpression Lysate Summary

Description

Aromatase Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC221358

Accession#: NM_031226

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
CYP19A1

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
57.7 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 Aromatase Overexpression Lysate

  • ARO1
  • ARO1EC 1.14.14.1
  • Aromatase
  • AROsubfamily XIX (aromatization of androgens)
  • CYAR
  • CYARMGC104309
  • CYP19
  • CYP19A1
  • CYPXIX
  • cytochrome P450, family 19, subfamily A, polypeptide 1
  • Cytochrome P-450AROM
  • EC 1.14.14.14
  • Estrogen synthase
  • microsomal monooxygenase

Background

Aromatase is a member of the cytochrome P450 superfamily, whose function is to produce estrogens by aromatizing androgens. The primary function of Aromatase is to convert androstenedione to estrone and testosterone to estradiol. This protein is also a key enzyme in steroidogenesis, specifically for the biosynthesis of estrogens. Because estrogens also promote certain cancers and other diseases, aromatase inhibitors are frequently used to treat such diseases. It also plays an important role in sexual differentiation, fertility, and carcinogenesis.

Aromatase is localized in the endoplasmic reticulum of the cell and its activity is regulated by tissue specific promoters that are in turn controlled by hormones, cytokines, and other factors. It can be found in many tissues reproductive tissues including gonads, brain, adipose tissue, placenta, blood vessels, skin, bone, endometrium as well as in tissue of endometriosis, uterine fibroids, breast cancer, and endometrial cancer.

Many environmental chemicals may influence aromatase activity and thereby disrupt endocrine function. The activity is increased by environmental factors such as age, obesity, insulin, gonadotropins, and alcohol but decreased by prolactin, anti-m llerian hormone, and smoking.

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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Aromatase - A Key Enzyme in the Biosynthesis of Estrogens
The enzyme, aromatase, belongs to cytochrome P450 family of monooxygenases known for their key role in drug catabolism and cholesterol/steroid synthesis. Aromatase uses a heme-group as a co-factor to catalyze the formation of aromatic C18 estrogens fr...  Read full blog post.

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Bioinformatics

Gene Symbol CYP19A1