Rab5a Overexpression Lysate

Name: Rab5a Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-15082

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-15082

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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 Rab5a Overexpression Lysate

Rab5a Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC203873

Accession#: NM_004162

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

23.5 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 Rab5a Overexpression Lysate

Western Blot: Rab5a Overexpression Lysate [NBL1-15082]

Western Blot: Rab5a Overexpression Lysate (Adult Normal) [NBL1-15082] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Rab5a.

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: Rab5a

Rab5 is a 24 kDa GTP-binding protein that regulates the fusion of plasma membrane -derived clathrin-coated vesicles with early endosomes , and homotypic fusion among early endosomes1. Localized to the cytoplasmic side of the plasma membrane, clathrin -coated vesicles and early endosomes, Rab5 appears to regulate vesicle fusion through a cycle of GDP/GTP e xchange and GTP hydrolysis. The different guanine nuc leotide binding states of Rab5 may affect its ability to associate or dissociate with membranes during endocytotic membrane traffic. The GTP-bound, or active, form of Rab5 associates with membrane s and regulates vesicle docking and fusion. Studies using a Rab5 mutant that hydrolyzed xanthosine 5 -triphosphate (XTP) indicated that nucleotide hydrolysis occurs even in the absence of membrane fusion. GTP hydrolysis by Rab5 may determine the frequency of membrane docking and fusion events.

Alternate Names

RAB5A, member RAS oncogene family, RAB5RAS-associated protein RAB5A, ras-related protein Rab-5A

Gene Symbol

RAB5A

Additional Rab5a Products

Product Documents for Rab5a Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Rab5a 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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