Product Specifications for Recombinant Human IL-32 alpha Protein, CF
E. coli-derived human IL-32 alpha protein Cys2-Lys131
>85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
18.8 kDa, reducing conditions
Measured by its ability to induce TNF-alpha secretion by RAW 264.7 mouse monocyte/macrophage cellsunder serum free conditions in the presence of muramyl dipeptide (MDP). Netea, M.G. et al. (2005) Proc. Nat. Acad. Sci. 102:16309. The ED50 for this effect is 2‑12 μg/mL.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS and DTT.
Reconstitute at 200 μg/mL in PBS.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-32 alpha
Interleukin 32 (IL-32) is an N-glycosylated cytokine that is up‑regulated by inflammatory stimulation in monocytes, NK cells, epithelial cells, and pancreatic myofibroblasts (1-5). It cooperates with these stimuli to promote the expression of other proinflammatory molecules such as TNF-alpha, IL-6, IL‑1 beta, IL-1 alpha, and CXCL8/IL-8 (5-7). The longest of several IL-32 splicing variants is the 20 ‑ 25 kDa gamma isoform which is also known as natural killer cell transcript 4 (NK4) (8, 9). The alpha isoform (IL-32 alpha ) lacks a portion of the putative signal peptide as well as 57 aa from the C-terminal region. IL-32 alpha is less potent than IL-32 beta, gamma, or δ at inducing the expression of proinflammatory molecules in peripheral blood mononuclear cells (PBMC) (8, 10). Neutrophil-derived Proteinase 3 (PR3) cleaves IL-32 alpha between Thr57 and Val58, a cleavage site that is retained in other IL-32 isoforms (11). The N-terminal fragment of PR3-cleaved IL-32 alpha shows increased potency at inducing CXCL2/MIP-2 and CXCL8 expression in PBMC relative to uncleaved IL-32 alpha (11, 12). IL-32 is highly expressed by colonic epithelial cells in inflammatory bowel disease and Crohn’s disease, rheumatoid arthritis synovium, and ductal epithelial cells in chronic pancreatitis and pancreatic cancer (5, 13 ‑ 15). IL-32 inhibits HIV-1 replication in vitro, and it is elevated in the serum of HIV-1 patients (16, 17).
Netea, M.G. et al. (2006) PloS Med. 3:e277.
Nold-Petry, C.A. et al. (2009) Proc. Natl. Acad. Sci. 106:3883.
Li, W. et al. (2009) Eur. J. Immunol. 39:1019.
Nishida, A. et al. (2008) Am. J. Physiol. Gastrointest. Liver Physiol. 294:G831.
Shoda, H. et al. (2006) Arthritis Res. Ther. 8:R166.
Netea, M.G. et al. (2005) Proc. Natl. Acad. Sci. 102:16309.
Hong, J. et al. (2010) Cytokine 49:171.
Kim, S.-H. et al. (2005) Immunity 22:131.
Dahl, C.A. et al. (1992) J. Immunol. 148:597.
Choi, J.-D. et al. (2009) Immunology 126:535.
Novick, D. et al. (2006) Proc. Natl. Acad. Sci. 103:3316.
Kim, S. et al. (2008) BMB Rep. 41:814.
Shioya, M. et al. (2007) Clin. Exp. Immunol. 149:480.
Joosten, L.A.B. et al. (2006) Proc. Natl. Acad. Sci. 103:3298.
Nishida, A. et al. (2009) J. Biol. Chem. 284:17868.
Rasool, S.T. et al. (2008) Immunol. Lett. 117:161.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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