Mineralocorticoid R/NR3C2 Overexpression Lysate

Western Blot: Mineralocorticoid Receptor Overexpression Lysate (Adult Normal) [NBP2-10715] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Mineralocorticoid Receptor.

• Reactivity: Hu
• Applications: WB

Mineralocorticoid R/NR3C2 Overexpression Lysate Summary

• Description:

  Mineralocorticoid R/NR3C2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220558
  
  Accession#: NM_000901
  
  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: NR3C2

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: 106.9 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 Mineralocorticoid R/NR3C2 Overexpression Lysate

• FLJ41052
• MGC133092
• Mineralocorticoid R
• mineralocorticoid receptor 1
• mineralocorticoid receptor delta
• MineralocorticoidR
• MLR
• MLRmineralocorticoid receptor
• MRMCRaldosterone receptor
• NR3C2
• NR3C2VIT
• Nuclear receptor subfamily 3 group C member 2
• nuclear receptor subfamily 3, group C, member 2

Background

Steroid receptors are ligand dependent, intracellular proteins that stimulate transcription of specific genes by binding to specific DNA sequences following activation by the appropriate hormone. Mineralocorticoids are a family of steroids, secreted by the adrenal cortex, necessary for the regulation of a number of metabolic processes including electrolyte regulation. These compounds exert their effect through their interaction with the mineralocorticoid receptor and that complex's subsequent association with DNA. Given the function of mineralocorticoids, it is not surprising to find that the kidney is a primary target organ for mineralocorticoids and that this organ has been shown to contain Mineralocorticoid Receptor.

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.

FAQs for Mineralocorticoid R/NR3C2 Lysate (NBP2-10715). (Showing 1 - 2 of 2 FAQ).

1. We need a positive control for the detection of mouse MR in Western blot and need to know whether the MR lysate (NBP2-10715) will recognize mouse MR?
   • NBP2-10715 is intended to be used as a positive control in western blot, however it is a human Mineralocorticoid Receptor Overexpression Lysate. Unfortunately, it looks like this is the only MR overexpression lysate we carry at this time.
2. Which antibody was used to detect the MR-positive control shown on the data sheet?
   • The western blot image has been generated using the anti-DYKDDDDK Epitope Tag antibody (NBP1-71705).

Bioinformatics

• Gene Symbol: NR3C2