MPRA Overexpression Lysate

Name: MPRA Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04977

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-04977

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for MPRA Overexpression Lysate

MPRA Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC204601

Accession#: NM_178422

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.

Product Specifications

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.

TMW

39.5 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.

Type

Overexpression

Scientific Data Images for MPRA Overexpression Lysate

Western Blot: MPRA Overexpression Lysate [NBP2-04977]

Western Blot: MPRA Overexpression Lysate (Adult Normal) [NBP2-04977] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MPRA.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: MPRA

The steroid progesterone induces the resumption of maturation in oocytes via a nongenomic pathway through binding to a novel, membrane progestin receptor (mPR). This pathway inhibits adenylyl cyclase and reduces intracellular cAMP, and also activates mitogen-activated protein kinase to effect signal transduction pathways. Three distinct groups, designated alpha, beta and gamma, comprise this gene family. mPRalpha, also designated progestin and adipoQ receptor family member VII (PAQR7), consists of an extracellular N-terminus, an intracellular C-terminus, and seven transmembrane domains. It is expressed in ovary, testis, placenta, uterus and bladder. mPRbeta, also designated progestin and adipoQ receptor family member VIII (PAQR8), consists of eight putative transmembrane regions and an intracellular N-terminus that contains a leucine-rich motif. It is a 354 amino acid protein with a molecular mass of about 40 kDa and is expressed in brain and spinal cord. Both mPRalpha and mPRbeta may be G protein-coupled receptors and may be involved in oocyte maturation.

Alternate Names

membrane progestin receptor alpha, mPR alpha, MPRA2310021M12Rik, MRPA, mSR, Progestin and adipoQ receptor family member 7, progestin and adipoQ receptor family member VIIPGLP

Gene Symbol

PAQR7

Additional MPRA Products

Product Documents for MPRA Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for MPRA Overexpression Lysate

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.

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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