CD-M6PR Overexpression Lysate

Name: CD-M6PR Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-10825

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10825

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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 CD-M6PR Overexpression Lysate

CD-M6PR Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC201277

Accession#: NM_002355

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

27.9 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.

Type

Overexpression

Scientific Data Images for CD-M6PR Overexpression Lysate

Western Blot: CD-M6PR Overexpression Lysate [NBP2-10825]

Western Blot: Mannose 6 Phosphate Receptor (Cation dependent) Overexpression Lysate (Adult Normal) [NBP2-10825] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Mannose 6 Phosphate Receptor (Cation dependent).

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: CD-M6PR

Lysosomal enzymes containing one or two mannose 6-phosphate (man6P) moieties are moved about in the cell by two distinct but interconnected cycles by means of 300 kDa cation independent mannose 6-phospate receptors (CI-MPR). The biosynthetic cycle refers to the MPR transport of newly synthesized lysosomal enzymes from the trans Golgi network to late endosomes or early lysosomes. The endocytic cycle transports extracellular lysosomal enzymes from the plasma membrane via clathrin-coated vesicles to early endosomes. The entire pool of MPRs cycles between these cellular compartments every 3 hours. The steady state distribution of MPR's is predominantly within late endosomes, fewer in the trans Golgi network and ~10 % at cell surface. In addition to its man6P binding activity, the MPR contains a separate binding site for the type II insulin-like growth factor and is capable of binding both man6P and IGF-II simultaneously. An ~240 kDa soluble, truncated form, representing the extracellular domain of the protein, has also been found circulating in serum and is capable of binding both ligands.

Long Name

Cation Dependent Mannose-6-Phosphate Receptor 46 kDa

Alternate Names

CD-MPR, CDM6PR, M6PR, MPR46, MPRD, SMPR

Gene Symbol

M6PR

Additional CD-M6PR Products

Product Documents for CD-M6PR Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for CD-M6PR 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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