Human CD30/TNFRSF8 Antibody

Catalog # Availability Size / Price Qty
AF229
AF229-SP
CD30/TNFRSF8 in Jurkat Human Cell Line.
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Human CD30/TNFRSF8 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD30 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross‑reactivity with recombinant mouse CD30 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD30
Phe19-Lys379
Accession # P28908
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human CD30/TNFRSF8 Fc Chimera (Catalog # 813-CD)
Agonist Activity
Measured by its ability to stimulate human IL-6 secretion by HDLM-2 human Hodgkin’s lymphoma cells. Gruss H.J. et al. (1996) Blood 87(6):2443.
The ED50 for this effect is typically 0.2 - 0.6 μg/mL.
 
Immunocytochemistry
5-15 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.

Data Example

Immunocytochemistry CD30/TNFRSF8 in Jurkat Human Cell Line. View Larger

CD30/TNFRSF8 in Jurkat Human Cell Line. CD30/TNFRSF8 was detected in immersion fixed Jurkat human acute T cell leukemia cell line using Goat Anti-Human CD30/TNFRSF8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF229) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell membrane and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: CD30/TNFRSF8

CD30, also known as Ki-1 antigen and TNFRSF8, is a 120 kDa type I transmembrane glycoprotein belonging to the TNF receptor superfamily (1, 2). Mature human CD30 consists of a 361 amino acid (aa) extracellular domain (ECD) with six cysteine-rich repeats, a 28 aa transmembrane segment, and a 188 aa cytoplasmic domain (3). In contrast, mouse and rat CD30 lack 90 aa of the ECD and contain only three cysteine-rich repeats. Within common regions of the ECD, human CD30 shares 53% and 49% aa sequence identity with mouse and rat CD30, respectively. Alternate splicing of human CD30 generates an isoform that includes only the C‑terminal 132 aa of the cytoplasmic domain. CD30 is normally expressed on antigen-stimulated Th cells and B cells (4 - 6). However, it is upregulated in Hodgkin’s disease (on Reed-Sternberg cells), other lymphomas, chronic inflammation, and autoimmunity (7). CD30 binds to CD30 Ligand/TNFSF8 which is expressed on activated Th cells, monocytes, granulocytes and medullary thymic epithelial cells (1, 5). CD30 signaling costimulates antigen-induced Th0 and Th2 proliferation and cytokine secretion but favors a Th2-biased immune response (8). In the absence of antigenic stimulation, it can still induce T cell expression of IL-13 (9). CD30 contributes to thymic negative selection by inducing the apoptotic cell death of CD4+CD8T cells (10, 11). In B cells, CD30 ligation promotes cellular proliferation and antibody production in addition to the expression of CXCR4, CCL3, and CCL5 (5, 12). An 85-90 kDa soluble form of CD30 is shed from the cell surface by TACE-mediated cleavage (13, 14). Soluble CD30 retains the ability to bind CD30 Ligand and functions as an inhibitor of normal CD30 signaling (15).

References
  1. Kennedy, M.K. et al. (2006) Immunology 118:143.
  2. Tarkowski, M. (2003) Curr. Opin. Hematol. 10:267.
  3. Durkop, H. et al. (1992) Cell 68:421.
  4. Hamann, D. et al. (1996) J. Immunol. 156:1387.
  5. Shanebeck, S.D. et al. (1995) Eur. J. Immunol. 25:2147.
  6. Gruss, H.-J. et al. (1994) Blood 83:2045.
  7. Oflzoglu E. et al. (2009) Adv. Exp. Med. Biol. 647:174.
  8. Del Prete, G. et al. (1995) J. Exp. Med. 182:1655.
  9. Harlin, H. et al. (2002) J. Immunol. 169:2451.
  10. Amakawa, R. et al. (1996) Cell 84:551.
  11. Chiarle, R. et al. (1999) J. Immunol. 163:194.
  12. Vinante, F. et al. (2002) Blood 99:52.
  13. Hansen, H.P. et al. (1995) Int. J. Cancer 63:750.
  14. Hansen, H.P. et al. (2000) J. Immunol. 165:6703.
  15. Hargreaves, P.G. and A. Al-Shamkhani (2002) Eur. J. Immunol. 32:163.
Entrez Gene IDs
943 (Human); 21941 (Mouse); 25069 (Rat)
Alternate Names
CD30 antigen; CD30; CD30KI-1; CD30L receptor; cytokine receptor CD30; D1S166EKi-1; Ki-1 antigen; Lymphocyte activation antigen CD30; TNFRSF8; tumor necrosis factor receptor superfamily member 8; tumor necrosis factor receptor superfamily, member 8

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