Skp2 Overexpression Lysate

Name: Skp2 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-15995

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-15995

Submit a review

Product Datasheet / COA / SDS

100% Guarantee

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

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Skp2 Overexpression Lysate

Skp2 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC214001

Accession#: NM_005983

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

47.6 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 Skp2 Overexpression Lysate

Western Blot: Skp2 Overexpression Lysate [NBL1-15995]

Western Blot: SKP2 Overexpression Lysate (Adult Normal) [NBL1-15995] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SKP2.

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: Skp2

The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation promoting factor). Cyclin A accumulates prior to cyclin B in the cell cycle, appears to be involved in control of S phase and has been shown to associate with cyclin dependent kinase 2 (Cdk2). In addition, cyclin A has been implicated in cell transformation and is found in complexes with E1A, transcription factors DP-1 and E2F, and retinoblastoma protein p110. Two cyclin A-Cdk2 complex binding proteins, Skp1 p19 and Skp2 p45, have been described. Although the Skps (S phase kinase-associated proteins) associate with the active cyclin A-Cdk2 complex, they do not exhibit any regulatory effects on the complex. Abolition of Skp2 p45 function by either microinjection of anti-p45 antibodies or addition of antisense oligonucleotides prevents entry into S phase of both normal and transformed cells.

Long Name

S Phase Kinase-associated Protein 2

Alternate Names

FBL1, FBXL1, p45skp2

Gene Symbol

SKP2

Additional Skp2 Products

Product Documents for Skp2 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Skp2 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.

Citations
Reviews