Human PBMC either (A) Activated with PMA (50 ng/ml) and Ca²⁺ Ionomycin (200 ng/ml) for 16 hours, or (B) untreated, were stained with Mouse Anti-Human CD30 Ligand/TNFSF8 Monoclonal Antibody (Catalog # MAB7741) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (F0102B) and Mouse anti-Human CD3 APC-conjugated Monoclonal Antibody (FAB100A). Quadrant markers were set based on control antibody staining (MAB003). Staining was performed using our Staining Membrane-associated Proteins protocol.

CD30 Ligand/TNFSF8 in Human Spleen.

CD30 Ligand/TNFSF8 was detected in immersion fixed paraffin-embedded sections of human spleen using Mouse Anti-Human CD30 Ligand/TNFSF8 Monoclonal Antibody (Catalog # MAB7741) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody VC001. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic CTS013. Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in splenic nodules. Staining was performed using our IHC Staining with our VisUCyte HRP Polymer Detection Reagents Protocol.

IL-6 Secretion Induced by CD30 Ligand/TNFSF8 and Neutralization by Human CD30 Ligand/TNFSF8 Antibody.

In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (10 µg/mL, MAB050), Recombinant Human CD30 Ligand/TNFSF8 (Catalog # Catalog # 1028-CL) stimulates IL-6 secretion in the HDLM human Hodgkin’s lymphoma cell line in a dose-dependent manner (orange line), as measured by the Human IL-6 Quantikine ELISA Kit (D6050). Under these conditions, IL-6 secretion elicited by Recombinant Human CD30 Ligand/TNFSF8 (1 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CD30 Ligand/TNFSF8 Monoclonal Antibody (Catalog # MAB7741). The ND₅₀ is typically 0.3-3 µg/mL.

Applications for Human CD30 Ligand/TNFSF8 Antibody

Application

Recommended Usage

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human PBMC activated with PMA (50 ng/ml) and Ca2+ Ionomycin (200 ng/ml) for 16 hours

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human spleen

Neutralization

Measured by its ability to neutralize CD30 Ligand/TNFSF8-induced IL‑6 secretion in the HDLM human Hodgkin's lymphoma cell line. The Neutralization Dose (ND₅₀) is typically 0.3-3 µg/mL in the presence of 1 µg/mL Recombinant Human CD30 Ligand/TNFSF8 and 10 µg/mL of a cross-linking antibody, Mouse Anti-His Tag Monoclonal Antibody.

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

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

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 Ligand/TNFSF8

CD30 ligand (CD30L)/TNFSF8 is a type II membrane protein belonging to the TNF superfamily. CD30L is expressed on the cell surface of activated T cells, B cells, and monocytes. The protein is also constitutively expressed on granulocytes and medullary thymic epithelial cells. The specific receptor for CD30L is CD30/TNFRSF8, a type I transmembrane glycoprotein belonging to the TNF receptor superfamily. CD30 was originally identified as a cell surface antigen of Hodgkin's and Reed-Sternberg cells using the monoclonal antibody Ki-1. CD30 is also expressed on different non-Hodgkin's lymphomas, virus-infected T and B cells, and on normal T and B cells after activation. Among T cells, CD30 is preferentially expressed on a subset of T cells producing Th2-type cytokines and on CD4⁺/CD8⁺ thymocytes that co-express CD45RO and IL-4 receptor. CD30 ligation by CD30L mediates pleiotropic effects including cell proliferation, activation, differentiation and cell death by apoptosis. CD30 can act as a co-stimulatory molecule in thymic negative selection and may also play a critical role in the pathophysiology of Hodgkin's disease and other CD30⁺ lymphomas. Human and mouse CD30 ligand cDNAs share 70% sequence homology.

References

Brunangelo, F. et al. (1995) Blood 85:1.
Duckett, C.S. et al. (1997) Mol. Cell. Biol. 17:1535.
Chiarle, R. et al. (1999) J. Immunol. 163:194.

Alternate Names

CD153, CD30L, TNFSF8

Entrez Gene IDs

944 (Human); 21949 (Mouse)

Gene Symbol

TNFSF8

UniProt

P32971

Additional CD30 Ligand/TNFSF8 Products

Product Documents for Human CD30 Ligand/TNFSF8 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human CD30 Ligand/TNFSF8 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human CD30 Ligand/TNFSF8 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB7741

Key Product Details

Species Reactivity

Validated:

Applications

Label

Antibody Source

View all CD30 Ligand/TNFSF8 Antibodies »

Product Summary for Human CD30 Ligand/TNFSF8 Antibody

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human CD30 Ligand/TNFSF8 Antibody

Detection of CD30 Ligand/TNFSF8 in Human PBMC by Flow Cytometry.