IL-32 Antibody (2525B)

R&D Systems | Catalog # MAB6769

Recombinant Monoclonal Antibody.
R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Rat

Applications

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2525B
View all IL-32 Primary Antibodies »

Product Specifications

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.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for IL-32 Antibody (2525B)

Detection of Human and Rat IL-32 antibody by Western Blot.

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.
IL-32 antibody in Human PBMCs by Immunocytochemistry (ICC).

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.

Applications for IL-32 Antibody (2525B)

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA

Western Blot

1 µg/mL
Sample: U937 human histiocytic lymphoma cell line and Nb2‑11 rat lymphoma cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 delta 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

  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.

Long Name

Interleukin 32

Alternate Names

IL32, NK4, TAIF

Entrez Gene IDs

9235 (Human)

Gene Symbol

IL32

UniProt

NP_001012649

Additional IL-32 Products

Product Documents for IL-32 Antibody (2525B)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for IL-32 Antibody (2525B)

For research use only

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Protocols

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