ASC2 Overexpression Lysate

Western Blot: ASC2 Overexpression Lysate (Adult Normal) [NBL1-15020] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ASC2.

• Reactivity: Hu
• Applications: WB

ASC2 Overexpression Lysate Summary

• Description:

  ASC2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211240
  
  Accession#: NM_152901
  
  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: PYDC1

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: 9.9 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 ASC2 Overexpression Lysate

• ASC2PAAD-only protein 1
• ASCI
• POP1pyrin domain-containing protein 1
• PYC1
• PYD (pyrin domain) containing 1
• pyrin domain containing 1
• pyrin-domain containing protein 1
• Pyrin-only protein 1

Background

ASC2/POP1 is a PAAD domain only protein originally identified in a bioinformatics screen aimed at understanding molecular apoptosis mechanisms (Pawtokski et al, 2001; reviewed in Fabiola et al, 2006, and Mariathasan and Vuvic, 2003.). ASC2/POP1 has high amino acid sequence homology with ASC (64%), hence it was originally termed ASC2. The PAAD (also known as PYRIN) domain is a conserved sequence motif identified in more than 35 human proteins with putative functions in apoptosis and inflammatory signaling pathways. PAAD was named after the protein families from which it was discovered: pyrin, AIM (absent-in-melanoma), ASC [apoptosis-associated speck-like protein containing a caspase recruitment domain (CARD], and death-domain (DD)-like. PAAD is thought to function as a protein-protein interaction domain, possibly coupling different signaling pathways such as apoptosis, inflammation and cancer. For example, PAADs are capable of homotypic interactions with themselves or other members of the PAAD family, suggesting that they participate in complex protein-interaction networks that link various signalling pathways. In humans, the gene encoding ASC2/POP1 is on chromosome 16p12.1, only 14 kbp away from the ASC locus. The close proximity of ASC2/POP1 to ASC as well as the high sequence homology between them suggest that the ASC2/POP1 and ASC genes arose by gene duplication. Studies have shown that ASC2/POP1 associates with ASC via PADD-PADD interactions and modulates ASC-mediated roles in apoptosis and inflammation. ASC2/POP1 may also have a role in modulating other multidomain PAAD-containing proteins. However, the physiological relevance of ASC2/POP1 remains to be fully elucidated. Human ASC2/POP1 is an 89 amino acid protein and migrates at ~10-12 kDa on SDS-PAGE gels (Stehlik et al, 2003).

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: PYDC1