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RAPGEF5 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-07464

Novus Biologicals, part of Bio-Techne
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NBP2-07464

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for RAPGEF5 Overexpression Lysate

RAPGEF5 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC218010

Accession#: NM_012294

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

84.5 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 RAPGEF5 Overexpression Lysate

Western Blot: RAPGEF5 Overexpression Lysate [NBP2-07464]

Western Blot: RAPGEF5 Overexpression Lysate [NBP2-07464]

Western Blot: RAPGEF5 Overexpression Lysate (Adult Normal) [NBP2-07464] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for RAPGEF5.

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: RAPGEF5

Members of the RAS (see HRAS; MIM 190020) subfamily of GTPases function in signal transduction as GTP/GDP-regulated switches that cycle between inactive GDP- and active GTP-bound states. Guanine nucleotide exchange factors (GEFs), such as RAPGEF5, serve as RAS activators by promoting acquisition of GTP to maintain the active GTP-bound state and are the key link between cell surface receptors and RAS activation (Rebhun et al., 2000 [PubMed 10934204]).[supplied by OMIM]

Alternate Names

GFRREPAC, Guanine nucleotide exchange factor for Rap1, KIAA0277M-Ras-regulated GEF, M-Ras-regulated Rap GEF, MRGEF, MR-GEFrap guanine nucleotide exchange factor 5, Rap guanine nucleotide exchange factor (GEF) 5, Related to Epac, Repac

Gene Symbol

RAPGEF5

Additional RAPGEF5 Products

Product Documents for RAPGEF5 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for RAPGEF5 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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