C5 lysate

Cynomolgus C5 HEK293 Overexpression Lysate

• Applications: Western Blot
• Synonyms: C5; Cynomolgus C5 HEK293 Overexpression Lysate; Cynomolgus C5 HEK293 Cell Lysate (WB positive control); complement component 5; C5 lysate

• Host: HEK293 Cells
• Form/Format: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.; 1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

• Applicable Applications for C5 lysate: Western Blot (WB)
• Species: Cynomolgus
• Sequence Information: A DNA sequence encoding the cynomolgus C5 (XP_005580972.1)(Gln19-Cys1676) was expressed with a polyhistidine tag at the C-terminus.
• Preparation Method: Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
• Recommend Usage: 1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.; 2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.; 3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
• Preparation and Storage: Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.

Related Product Information for C5 lysate

Description: This Cynomolgus C5 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of C5 protein from the overexpression lysate was verified.

Background: C5a is a protein fragment released from complement component C5. This 74 amino acid peptide in humans is generated by the cleavage of C5a convertase on the C5 alpha-chain during the classical, alternative, and lectin pathways of complement activation. The structure of C5a includes a core region consisting of four, anti-parallel alpha-helices held together by three disulfide linkages and a structured C-terminal tail, and C5a is rapidly metabolised by carboxypeptidase B to a 73 amino acid low activity form, C5a des-Arg. C5a is an extremely potent proinflammatory mediator, as well as a potent chemotactic factor for neutrophils and other leukocytes. It causes histamine release, increases in vascular permeability, induces several cytokines production from leukocytes, enhances neutrophil-endothelial cell adhesion, and augments the humoral and cell-mediated immune response. C5a is quickly metabolised by carboxypeptidases, forming the less potent C5adesArg. Acting via a classical G protein-coupled receptor, CD88, C5a and C5adesArg exert a number of effects essential to the innate immune response, while their actions at the more recently discovered non-G protein-coupled receptor, C5L2 (or GPR77), remain unclear. The widespread expression of C5a receptors throughout the body allows C5a to elicit a broad range of effects. Thus, C5a has been found to be a significant pathogenic driver in a number of immuno-inflammatory diseases, making C5a inhibition an attractive therapeutic strategy. C5a is a strong chemoattractant and is involved in the recruitment of inflammatory cells such as neutrophils, eosinophils, monocytes, and T lymphocytes, in activation of phagocytic cells and release of granule-based enzymes and generation of oxidants, all of which may contribute to innate immune functions or tissue damage. Accordingly, the anaphylatoxin C5a is implicated in a variety of diseases such as rheumatoid arthritis, systemic lupus erythematosus, reperfusion injury, Alzheimer's disease, and sepsis.

Product Categories/Family for C5 lysate

Complement Activation Pathways

References

Guo RF, et al.. (2005) Role of C5a in inflammatory responses. Annu Rev Immunol. 23: 821-52.Guo RF, et al. (2006) C5a, a therapeutic target in sepsis. Recent Pat Antiinfect Drug Discov. 1(1): 57-65.Manthey HD, et al. (2009) Complement component 5a (C5a). Int J Biochem Cell Biol. 41(11): 2114-7.