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The specific activity is determined by proliferation assay using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Human CD40-Ligand
Figure 1
Human CD40-Ligand activity assay:
The specific activity is determined by proliferation assay using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Human CD40-Ligand
Figure 2
Human CD40-Ligand activity assay:
The specific activity is determined by
activation assay
using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Mass spectrometry analysis (ESI-MS) of Human CD40-Ligand, premium grade. The major peak corresponds to the calculated molecular mass of 23034 Da. The minor peaks correspond to gluconoylation derivatives as described in Geoghegan, K. F.
et al.
(1999).
Human CD40-Ligand
Figure 5
Mass spectrometry analysis (ESI-MS) of Human CD40-Ligand, premium grade. The major peak corresponds to the calculated molecular mass of 23034 Da. The minor peaks correspond to gluconoylation derivatives as described in Geoghegan, K. F.
et al.
(1999).
Specifications for Human CD40-Ligand
Overview
Human CD40-Ligand is a recombinant protein optimized for use in cell culture, differentiation studies, and functional assays.
Applications
Human CD40-Ligand can be used for a variety of applications, including:
Stimulation of B cell activation and proliferation.
Stimulation of dendritic cell maturation.
Induction of cytokine production in peripheral blood monocytes and T cells.
Alternative names
TRAP, CD154, TNFSF5
Detailed product information
Background information
CD40-Ligand, also known as CD40L, CD154, TRAP, or TNFSF5 is a member of the TNF superfamily transiently expressed on the surface of activated CD4
+
T lymphocytes. It is either expressed membrane-bound or proteolytically released as a soluble form which comprises 2/3 of the extracellular domain. Its receptor CD40 is found on antigen-presenting cells such as B cells, dendritic cells, and macrophages. CD40L-CD40 interaction is important in T cell-APC (antigen-presenting cell) interaction and is e.g. involved in B cell differentiation and proliferation, isotype class-switching, and protection of B cells from apoptosis.
Biological activity
Proliferation of CD19
+
B cells
premium grade: ≥ 4×
10
3
U/mg
We measure the biological activity of each batch of MACS Premium-Grade Cytokines and state the results in the Certificate of Analysis (CoA). Based on the lot-specific activity, exact doses of active cytokine can be applied to cell culture experiments. This allows for reproducible cell culture conditions without the need for time-consuming lot-to-lot testing.
Quality description
Premium-grade
cytokines offer the convenience of high and well-defined biological activities and allow exact unit dosing for demanding applications. The biological activity is determined after lyophilization and reconstitution, and normalized to WHO/NIBSC standards whenever available. In general, endotoxin levels are <0.01 ng/μg (<0.1 EU/μg), and purities are >97%. Lot-specific activities are stated in the Certificate of Analysis (www.miltenyibiotec.com/certificates).
to search for Certificates of Analysis (CoA) by lot number.
References for Human CD40-Ligand
Publications
Spriggs, M. K. et al. (1992) Recombinant human CD40 ligand stimulates B cell proliferation and immunoglobulin E secretion. J. Exp. Med. 176: 1543-1550
Hou, G. et al. (2021) SLE non-coding genetic risk variant determines the epigenetic dysfunction of an immune cell specific enhancer that controls disease-critical microRNA expression. Nat Commun. 12(1): 135
Kuen, J. et al. (2017) Pancreatic cancer cell/fibroblast co-culture induces M2 like macrophages that influence therapeutic response in a 3D model. PLoS One 12(7): e0182039
De Keersmaecker, B. et al. (2020) TriMix and tumor antigen mRNA electroporated dendritic cell vaccination plus ipilimumab: link between T-cell activation and clinical responses in advanced melanoma. J Immunother Cancer 8(1): e000329
Johnson, M. J. et al. (2018) Transcriptional profiles of human islet and exocrine endothelial cells in subjects with or without impaired glucose metabolism. Sci Rep 8(1): 12144
Horns, F. et al. (2016) Lineage tracing of human B cells reveals the in vivo landscape of human antibody class switching. Elife 5: e16578
Koliha, N. et al. (2016) Melanoma affects the composition of blood cell-derived extracellular vesicles. Front Immunol 7: 282
Monaghan, T. M. et al. (2013) Circulating antibody and memory B-Cell responses to C. difficile toxins A and B in patients with C. difficile-associated diarrhoea, inflammatory bowel disease and cystic fibrosis PLoS One 8(9): e74452
The specific activity is determined by proliferation assay using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Human CD40-Ligand
Figure 1
Human CD40-Ligand activity assay:
The specific activity is determined by proliferation assay using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Human CD40-Ligand
Figure 2
Human CD40-Ligand activity assay:
The specific activity is determined by
activation assay
using enriched CD19
+
B cells in the presence of the cross-linking antibody and of 50 IU/mL Interleukin 4. Activity of Human CD40-Ligand, premium grade (red line) was compared to another commercially available product (black line).
Mass spectrometry analysis (ESI-MS) of Human CD40-Ligand, premium grade. The major peak corresponds to the calculated molecular mass of 23034 Da. The minor peaks correspond to gluconoylation derivatives as described in Geoghegan, K. F.
et al.
(1999).
Human CD40-Ligand
Figure 5
Mass spectrometry analysis (ESI-MS) of Human CD40-Ligand, premium grade. The major peak corresponds to the calculated molecular mass of 23034 Da. The minor peaks correspond to gluconoylation derivatives as described in Geoghegan, K. F.
Human CD40-Ligand is a recombinant protein optimized for use in cell culture, differentiation studies, and functional assays.
Applications
Human CD40-Ligand can be used for a variety of applications, including:
Stimulation of B cell activation and proliferation.
Stimulation of dendritic cell maturation.
Induction of cytokine production in peripheral blood monocytes and T cells.
Alternative names
TRAP, CD154, TNFSF5
Detailed product information
Background information
CD40-Ligand, also known as CD40L, CD154, TRAP, or TNFSF5 is a member of the TNF superfamily transiently expressed on the surface of activated CD4
+
T lymphocytes. It is either expressed membrane-bound or proteolytically released as a soluble form which comprises 2/3 of the extracellular domain. Its receptor CD40 is found on antigen-presenting cells such as B cells, dendritic cells, and macrophages. CD40L-CD40 interaction is important in T cell-APC (antigen-presenting cell) interaction and is e.g. involved in B cell differentiation and proliferation, isotype class-switching, and protection of B cells from apoptosis.
Biological activity
Proliferation of CD19
+
B cells
premium grade: ≥ 4×
10
3
U/mg
We measure the biological activity of each batch of MACS Premium-Grade Cytokines and state the results in the Certificate of Analysis (CoA). Based on the lot-specific activity, exact doses of active cytokine can be applied to cell culture experiments. This allows for reproducible cell culture conditions without the need for time-consuming lot-to-lot testing.
Quality description
Premium-grade
cytokines offer the convenience of high and well-defined biological activities and allow exact unit dosing for demanding applications. The biological activity is determined after lyophilization and reconstitution, and normalized to WHO/NIBSC standards whenever available. In general, endotoxin levels are <0.01 ng/μg (<0.1 EU/μg), and purities are >97%. Lot-specific activities are stated in the Certificate of Analysis (www.miltenyibiotec.com/certificates).
Spriggs, M. K. et al. (1992) Recombinant human CD40 ligand stimulates B cell proliferation and immunoglobulin E secretion. J. Exp. Med. 176: 1543-1550
Hou, G. et al. (2021) SLE non-coding genetic risk variant determines the epigenetic dysfunction of an immune cell specific enhancer that controls disease-critical microRNA expression. Nat Commun. 12(1): 135
Kuen, J. et al. (2017) Pancreatic cancer cell/fibroblast co-culture induces M2 like macrophages that influence therapeutic response in a 3D model. PLoS One 12(7): e0182039
De Keersmaecker, B. et al. (2020) TriMix and tumor antigen mRNA electroporated dendritic cell vaccination plus ipilimumab: link between T-cell activation and clinical responses in advanced melanoma. J Immunother Cancer 8(1): e000329
Johnson, M. J. et al. (2018) Transcriptional profiles of human islet and exocrine endothelial cells in subjects with or without impaired glucose metabolism. Sci Rep 8(1): 12144
Horns, F. et al. (2016) Lineage tracing of human B cells reveals the in vivo landscape of human antibody class switching. Elife 5: e16578
Koliha, N. et al. (2016) Melanoma affects the composition of blood cell-derived extracellular vesicles. Front Immunol 7: 282
Monaghan, T. M. et al. (2013) Circulating antibody and memory B-Cell responses to C. difficile toxins A and B in patients with C. difficile-associated diarrhoea, inflammatory bowel disease and cystic fibrosis PLoS One 8(9): e74452
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