IL-32 Antibody (2525B) [Unconjugated]

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.

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.

• Reactivity: Hu, Rt
• Applications: WB, ICC/IF
• Clone: 2525B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

IL-32 Antibody (2525B) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E. coli-derived human IL‑32 beta protein
  Met1-Lys188
  Accession # NP_001012649
• 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: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-32 Antibody (2525B) [Unconjugated]

• 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

Background

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).

1. Netea, M.G. et al. (2006) PloS Med. 3:e277.
2. Nold-Petry, C.A. et al. (2009) Proc. Natl. Acad. Sci. USA 106:3883.
3. Li, W. et al. (2009) Eur. J. Immunol. 39:1019.
4. Nishida, A. et al. (2008) Am. J. Physiol. Gastrointest. Liver Physiol. 294:G831.
5. Shoda, H. et al. (2006) Arthritis Res. Ther. 8:R166.
6. Netea, M.G. et al. (2005) Proc. Natl. Acad. Sci. USA 102:16309.
7. Hong, J. et al. (2010) Cytokine 49:171.
8. Kim, S-H. et al. (2005) Immunity 22:131.
9. Dahl, C.A. et al. (1992) J. Immunol. 148:597.
10. Choi, J-D. et al. (2009) Immunology 126:535.
11. Novick, D. et al. (2006) Proc. Natl. Acad. Sci. USA 103:3316.
12. Kim, S. et al. (2008) BMB Rep. 41:814.
13. Shioya, M. et al. (2007) Clin. Exp. Immunol. 149:480.
14. Joosten, L.A.B. et al. (2006) Proc. Natl. Acad. Sci. USA 103:3298.
15. Nishida, A. et al. (2009) J. Biol. Chem. 284:17868.
16. Rasool, S.T. et al. (2008) Immunol. Lett. 117:161.
17. Nold, M.F. et al. (2008) J. Immunol. 181:557.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • NP_001012649()