KRAS Overexpression Lysate

Western Blot: KRAS Overexpression Lysate (Adult Normal) [NBL1-12376] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KRAS.

• Reactivity: Hu
• Applications: WB

KRAS Overexpression Lysate Summary

• Description:

  KRAS Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC201958
  
  Accession#: NM_004985
  
  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: KRAS

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: 21.2 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 KRAS Overexpression Lysate

• cellular c-Ki-ras2 proto-oncogene
• c-Ki-ras
• C-K-RAS
• GTPase KRas
• KI-RAS
• Kirsten rat sarcoma-2 viral (v-Ki-ras2) oncogene homolog
• K-Ras 2
• K-ras p21 protein
• KRAS1
• K-RAS2A
• K-RAS2B
• KRAS2NS3
• K-RAS4A
• K-RAS4B
• NS
• oncogene KRAS2
• PR310 c-K-ras oncogene
• RASK2c-Kirsten-ras protein
• transforming protein p21
• v-Ki-ras2 Kirsten rat sarcoma 2 viral oncogene homolog
• v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog

Background

This gene, a Kirsten ras oncogene homolog from the mammalian ras gene family, encodes a protein that is a member of the small GTPase superfamily. A single amino acid substitution is responsible for an activating mutation. The transforming protein that results is implicated in various malignancies, including lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas and colorectal carcinoma. Alternative splicing leads to variants encoding two isoforms that differ in the C-terminal region. [provided by RefSeq]. Transcript Variant: This variant (b) is composed of five exons and lacks exon 4a which the longer transcript variant (a) includes. This predominant variant (b) has a cds that terminates in exon 4b and encodes isoform b. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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.

Bioinformatics

• Gene Symbol: KRAS