Human CD30/TNFRSF8 Antibody

Catalog #: AF229 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF229
AF229-SP

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All CD30/TNFRSF8 Antibodies

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

Human CD30/TNFRSF8 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CD30 in direct ELISAs and Western blots.

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 either lyophilized or 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)

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human CD30/TNFRSF8 Monoclonal Antibody (Catalog # MAB2291).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human CD30/TNFRSF8 DuoSet ELISA Kit (Catalog # DY6126-05) for convenient development of a sandwich ELISA.

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.

Scientific Data

Immunocytochemistry

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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; 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.

ELISA

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Human CD30/TNFRSF8 ELISA Standard Curve. Recombinant Human CD30/TNFRSF8 protein was serially diluted 2-fold and captured by Mouse Anti-Human CD30/TNFRSF8 Monoclonal Antibody (MAB2291) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Human CD30/TNFRSF8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF229) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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⁺CD8⁺T 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

Kennedy, M.K. et al. (2006) Immunology 118:143.
Tarkowski, M. (2003) Curr. Opin. Hematol. 10:267.
Durkop, H. et al. (1992) Cell 68:421.
Hamann, D. et al. (1996) J. Immunol. 156:1387.
Shanebeck, S.D. et al. (1995) Eur. J. Immunol. 25:2147.
Gruss, H.-J. et al. (1994) Blood 83:2045.
Oflzoglu E. et al. (2009) Adv. Exp. Med. Biol. 647:174.
Del Prete, G. et al. (1995) J. Exp. Med. 182:1655.
Harlin, H. et al. (2002) J. Immunol. 169:2451.
Amakawa, R. et al. (1996) Cell 84:551.
Chiarle, R. et al. (1999) J. Immunol. 163:194.
Vinante, F. et al. (2002) Blood 99:52.
Hansen, H.P. et al. (1995) Int. J. Cancer 63:750.
Hansen, H.P. et al. (2000) J. Immunol. 165:6703.
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