CDC42 Overexpression Lysate

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Western Blot: CDC42 Overexpression Lysate (Adult Normal) [NBP2-05863] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CDC42.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB
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CDC42 Overexpression Lysate Summary

Description

CDC42 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216559

Accession#: NM_044472

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.
Gene
CDC42

Applications/Dilutions

Dilutions
  • Western Blot
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.
Theoretical MW
21.1 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.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

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.

Alternate Names for CDC42 Overexpression Lysate

  • CDC42
  • CDC42Hs
  • cell division control protein 42 homolog
  • cell division cycle 42 (GTP binding protein, 25kDa)
  • cell division cycle 42 (GTP-binding protein, 25kD)
  • dJ224A6.1.1 (cell division cycle 42 (GTP-binding protein, 25kD))
  • dJ224A6.1.2 (cell division cycle 42 (GTP-binding protein, 25kD))
  • G25K GTP-binding protein
  • G25K
  • growth-regulating protein
  • GTP-binding protein, 25kD
  • small GTP binding protein CDC42

Background

CDC42 is a small GTPase of the Rho subfamily, which regulates signaling pathways that control diverse cellular functions including cell morphology, migration, endocytosis and cell cycle progression. This protein is highly similar to Saccharomyces cerevisiae Cdc42, and is able to complement the yeast cdc42 1 mutant. The product of oncogene Dbl was reported to specifically catalyze the dissociation of GDP from this protein. This protein could regulate actin polymerization through its direct binding to Neural Wiskott Aldrich syndrome protein (NWASP), which subsequently activates Arp2/3 complex. Alternative splicing of this gene results in at least two transcript variants.

Limitations

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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Bioinformatics

Gene Symbol CDC42