Human CD30 Ligand/TNFSF8 Antibody

Catalog #: MAB7741 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7741-100
MAB7741-SP

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

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Human CD30 Ligand/TNFSF8 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CD30 LIgand/TNFSF8 in direct ELISAs.

Source

Monoclonal Mouse IgG_2A Clone # 116632

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived human CD30 Ligand/TNFSF8.
Gln63-Asp234
Accession # P32971

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

Flow Cytometry

0.25 µg/10⁶ cells

Human PBMC activated with PMA (50 ng/ml) and Ca2+ Ionomycin (200 ng/ml) for 16 hours

Immunohistochemistry

5-25 µg/mL

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 should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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

Immunohistochemistry

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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.

Neutralization

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

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.

Entrez Gene IDs

944 (Human); 21949 (Mouse)

Alternate Names

CD153 antigen; CD153; CD30 antigen ligand; CD30 Ligand; CD30L; CD30-L; CD30LCD30 ligand; CD30LGMGC138144; TNFSF8; tumor necrosis factor (ligand) superfamily, member 8; tumor necrosis factor ligand superfamily member 8