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Human Interleukin-32 / IL-32 (isoform alpha) HEK293 Cell Lysate (WB positive control)

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Human IL32 Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Human Cells
Product Description:Human Cell lysate that Human Interleukin-32 / IL-32 (isoform alpha) transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the human IL32 isoform alpha (IL32A) (NP_001012651.1) precursor (Met 1-Lys 131) was expressed with a polyhistidine tag at the C-terminus.
Predicted N Terminal:Met 1
Molecule Mass:The recombinant human IL32A comprises 137 amino acids with a predicted molecular mass of 15.7 kDa. As a result of glycosylation, it migrates as an approximately 20 kDa band in SDS-PAGE under reducing conditions.
Human IL32 Transfected / Overexpression Cell Lysate Usage Guide
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 (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
IL32 Background

IL-32 is a recently discovered cytokine that induces various proinflammatory cytokines (TNF-alpha, IL-1beta, IL-6) and chemokines in both human and mouse cells through the NF-kappaB and p38 MAPK inflammatory signal pathways. It is regulated robustly by other major proinflammatory cytokines, and is crucial to inflammation and immune responses. Four of the IL-32 isoforms (alpha, beta, gamma and delta) are the most representative IL-32 transcripts, and gamma isoform of IL-32 is the most active, although all isoforms are biologically active. IL-32, a cytokine produced mainly by T, natural killer, and epithelial cells induces significant amounts of TNFalpha and MIP-2 and increases the production of both cytokines in a dose-dependent manner. IL-32 has been implicated in inflammatory disorders, mycobacterium tuberculosis infections, inflammatory bowel disease, and influenza A virus infection, as well as in some autoimmune diseases, such as rheumatoid arthritis, ulcerative colitis and in human stomach cancer, human lung cancer and breast cancer tissues. Thus, IL-32 expression might be valuable as a biomarker for cancer.

Human IL32 References
  • Felaco P, et al. (2009) IL-32: a newly-discovered proinflammatory cytokine. J Biol Regul Homeost Agents. 23(3): 141-7.
  • Kobayashi H, et al. (2009) Molecular characterization of IL-32 in human endothelial cells. Cytokine. 46(3): 351-8.
  • Meyer N, et al. (2010) IL-32 is expressed by human primary keratinocytes and modulates keratinocyte apoptosis in atopic dermatitis. J Allergy Clin Immunol. 125(4): 858-865.
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    Catalog: 11064-H08HL-300
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