This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

Name: Recombinant Human CD30 Ligand/TNFSF8 Protein, CF
Brand: R&D Systems
SKU: 1028-CL

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Human CD30 Ligand/TNFSF8 Protein, CF

Catalog # 1028-CL

Citations (7)

Product Datasheet / COA / SDS

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Key Product Details

Source:	NS0
Accession #:	P32971
Applications:	Bioactivity

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Product Specifications

Source

Mouse myeloma cell line, NS0-derived human CD30 Ligand/TNFSF8 protein

MHHHHHHHHHH	GGGSGGGSGGGS	IEGR	Human CD30 Ligand (Gln63-Asp234) Accession # P32971
N-terminus			C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin level

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

N-terminal sequence Analysis

Met

Predicted Molecular Mass

22 kDa

SDS-PAGE

30-40 kDa, reducing conditions

Activity

Measured by its ability to stimulate IL-6 secretion by HDLM human Hodgkin’s lymphoma cells. Duckett, C.S. et al. (1997) Mol. Cell. Biol. 17:1535. The ED₅₀ for this effect is 8-48 ng/mL in the presence of 10 µg/mL of a cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1028-CL

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Shipping	The product is shipped at ambient temperature. Upon receipt, store it 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, -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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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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.

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

Citations for Recombinant Human CD30 Ligand/TNFSF8 Protein, CF (7)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human CD30 Ligand/TNFSF8 Protein, CF include:

Species: Human
Sample Types: Whole Cells
Applications: Cell Culture

M Yeste-Vela et al. (2019), The potential of brentuximab vedotin, alone or in combination with current clinical therapies, in the treatment of testicular germ cell tumors Am J Cancer Res, 9(5):855-871.
PMID: 31218098
Species: Human
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance (SPR

AW Clarke et al. (2018), An anti-TL1A antibody for the treatment of asthma and inflammatory bowel disease MAbs, 0(0):1-43.
PMID: 29436901
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Wilson TR et al. (2012), Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature, 487(7408):505-9.
PMID: 22763448
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Suzuki O et al. (2012), Galectin-1-mediated cell death is increased by CD30-induced signaling in anaplastic large cell lymphoma cells but not in Hodgkin lymphoma cells. Lab. Invest., 92(2):191-9.
PMID: 21986812
Species: Human

Applications: Surface Plasmon Resonance

Lobito AA et al. (2011), Murine insulin growth factor-like (IGFL) and human IGFL1 proteins are induced in inflammatory skin conditions and bind to a novel tumor necrosis factor receptor family member, IGFLR1. J. Biol. Chem., 286(21):18969-81.
PMID: 21454693
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Simhadri VL et al. (2011), A novel role for reciprocal CD30-CD30L signaling in the cross-talk between natural killer and dendritic cells. Biol. Chem., 393(1):101-6.
PMID: 22628304
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Schwertassek U et al. (2007), Selective redox regulation of cytokine receptor signaling by extracellular thioredoxin-1. EMBO J., 26(13):3086-97.
PMID: 17557078

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