NOD1 Overexpression Lysate Summary
Description |
NOD1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC203759
Accession#: NM_006092
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 |
NOD1 |
Applications/Dilutions
Dilutions |
|
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 |
107.5 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
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 NOD1 Overexpression Lysate
Background
Innate immunity is present in all animals and is the common mode of defense against microorganisms. It detects microorganisms by specific proteins called pattern-recognition molecules (PRMs) (reviewed in Werts et al. 2006). There is a limited set of PRMs in each animal genome, and it has been postulated that the PRMs were evolutionarily selected to detect conserved components or motifs of microorganisms called pathogen-associated molecular patterns (PAMPs). PAMPs are found in a wide range of microorganisms and recognition of PAMPs by PRMs activates inflammatory signaling pathways, thereby stimulating an immune response. NOD (nucleotide-binding oligomerization domain) proteins are a family of cytosolic proteins which have been implicated in innate recognition of bacteria, the induction of inflammatory responses, and the regulation of caspase activation and apoptosis. NOD1/CARD4 (caspase-recruitment domain 4 gene) is a PRM that recognizes specific peptidoglycan (PGN) components of bacterial cell walls (reviewed in Strober et al. 2006, and Inohara et al. 2003). NOD1 is expressed by cell types that are exposed to PGN under physiological conditions including antigen-presenting cells (APCs) such as macrophages and dendritic cells, and epithelial cells (reviewed in Strober et al. 2006). NOD1 is thought to play a role in the pathogenesis of human gastrointestinal disease and is associated with inflammatory bowel diseases and asthma. It is involved in host defense against Helicobacter pylori infection of the gastric mucosa, a chronic infection that can lead to peptic ulcers and gastric cancer. This antibody recognizes NOD1/CARD4. Human NOD1/CARD4 is a 953 amino acid protein.
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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