Human/Rat IL-32 Antibody

Catalog #: MAB6769 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB6769-100
MAB6769-SP

Detection of Human and Rat IL‑32 by Western Blot.

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All IL-32 Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Rat IL-32 Antibody Summary

Species Reactivity

Human, Rat

Specificity

Detects human IL-32 beta and other IL-32 isoforms in direct ELISAs. In direct ELISAs, 95% cross-reactivity with recombinant human (rh) IL-32 alpha and 45% cross-reactivity with rhIL-32 gamma is detected.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2525B

Purification

Protein A or G purified from cell culture supernatant

Immunogen

E. coli-derived human IL‑32 beta protein
Met1-Lys188
Accession # NP_001012649

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunocytochemistry

3-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human and Rat IL‑32 by Western Blot. Western blot shows lysates of Nb2-11 rat lymphoma cell line and U937 human histiocytic lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Rat IL-32 Monoclonal Antibody (Catalog # MAB6769) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for IL-32 at approximately 25 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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IL-32 in Human PBMCs. IL-32 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated or untreated with calcium ionomycin and PMA using Rabbit Anti-Human/Rat IL-32 Monoclonal Antibody (Catalog # MAB6769) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-32

Interleukin 32 (IL-32) is an N-glycosylated cytokine that is upregulated 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).

References

Netea, M.G. et al. (2006) PloS Med. 3:e277.
Nold-Petry, C.A. et al. (2009) Proc. Natl. Acad. Sci. USA 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. USA 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. USA 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. USA 103:3298.
Nishida, A. et al. (2009) J. Biol. Chem. 284:17868.
Rasool, S.T. et al. (2008) Immunol. Lett. 117:161.
Nold, M.F. et al. (2008) J. Immunol. 181:557.

Long Name

Interleukin 32

Entrez Gene IDs

9235 (Human)

Alternate Names

IL32; IL-32; IL-32alpha; IL-32beta; interleukin 32; interleukin-32 theta; interleukin-32; natural killer cell transcript 4; Natural killer cells protein 4; NK4; NK4IL-32delta; TAIF; TAIFa; TAIFb; TAIFc; TAIFd; TAIFIL-32gamma; Tumor necrosis factor alpha-inducing factor