Human CD30 Ligand/TNFSF8 Antibody

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 IgG2A 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 Detection of CD30 Ligand/TNFSF8 antibody in Human PBMC antibody by Flow Cytometry. View Larger

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 CD30 Ligand/TNFSF8 antibody in Human Spleen by Immunohistochemistry (IHC-P). View Larger

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 View Larger

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

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 0.5 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 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
  1. Brunangelo, F. et al. (1995) Blood 85:1.
  2. Duckett, C.S. et al. (1997) Mol. Cell. Biol. 17:1535.
  3. 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

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Isotype Controls

Mouse IgG2A Isotype Control

Reconstitution Buffers

Secondary Antibodies

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