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VPS34 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-14416

Novus Biologicals, part of Bio-Techne
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NBL1-14416

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for VPS34 Overexpression Lysate

VPS34 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC205317

Accession#: NM_002647

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

101.4 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 VPS34 Overexpression Lysate

Western Blot: VPS34 Overexpression Lysate [NBL1-14416]

Western Blot: VPS34 Overexpression Lysate [NBL1-14416]

Western Blot: VPS34 Overexpression Lysate (Adult Normal) [NBL1-14416] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for VPS34.

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: VPS34/PIK3C3

VPS34, also known as phosphoinositide-3-kinase p100 subunit or PIK3C3 (phosphoinositide-3-kinase, class III), is a member of the PI3/PI4-kinase family. It is ubiquitously expressed with predominant expression in skeletal muscle and is believed to participate in endosome to lysosome transport, multivesicular body formation, autophagy and retrograde endosome to Golgi transport. VPS34 is the catalytic subunit of class III PI3Ks and forms a heterodimer with p150, a regulatory subunit of class 3 PI3Ks. It exclusively phosphorylates phosphatidylinositol to produce PtdIns3P. The activity of VPS34 is enhanced by manganese and can be regulated by nutrients, suggesting an important role of VPS34 in the regulation of mTOR protein synthesis. VPS34 also forms a complex with Beclin1 that promotes autophagy and tumor supression.

Alternate Names

PI 3-Kinase C3, PI3K type 3, PIK3C3

Gene Symbol

PIK3C3

Additional VPS34/PIK3C3 Products

Product Documents for VPS34 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 VPS34 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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