INCA Overexpression Lysate

Western Blot: INCA Overexpression Lysate (Adult Normal) [NBP2-10988] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for INCA.

• Reactivity: Hu
• Applications: WB

INCA Overexpression Lysate Summary

• Description:

  INCA Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC224934
  
  Accession#: NM_001007232
  
  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: CARD17

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: 11.7 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 INCA Overexpression Lysate

• caspase recruitment domain family, member 17
• caspase recruitment domain-containing protein 17
• Caspase-1 inhibitor INCA
• INCAInhibitory CARD
• inhibitory caspase recruitment domain (CARD) protein
• Inhibitory caspase recruitment domain protein

Background

INCA, also known as CAR17 or Caspase recruitment domain-containing protein 17, is a 110 amino acid protein that is 12 kDa; cytoplasm located; regulates procaspase-1/CASP1 activation involved in the regulation of the proteolytic maturation of pro-IL-1beta/IL1B and its release during inflammation; and acts as an inhibitor of the release of IL1B in response to LPS in monocytes. Current research is being pursued on this protein involvement in several diseases and disorders including cancer, familial mediterranean fever, shigellosis, cowpox, neurodegenerative disease, vascular disease, mevalonic aciduria, tularemia, dermatitis, lung disease, legionnaires' disease, and arthritis. This protein has been linked to the proteolysis and regulation of apoptotic process pathways where it interacts with CASP1, CARD16, and CARD18 proteins.

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: CARD17