High activity fusion construct of human soluble CD40L (ab168061)

Overview

Description

  • Nature
    Recombinant
  • Source
    CHO cells
  • Amino Acid Sequence
    • Accession
    • Species
      Human
    • Sequence
      GDQNPQIAAHVISEASSKTTSVLQWAEKGYYTMSNNLVTLENGKQLTVKR QGLYYIYAQVTFCSNREASSQAPFIASLCLKSPGRFERILLRAANTHSSA KPCGQQSIHLGGVFELQPGASVFVNVTDPSQVSHGTGFTSFGLLKL
    • Molecular weight
      38 kDa
    • Amino acids
      116 to 261
    • Tags
      DDDDK tag N-Terminus
    • Additional sequence information
      The extracellular domain of human CD40L (aa 116-261) fused to N-ter to mouse collagen domain of Adiponectin (aa 18-111) & a FLAG®-tag. SwissProt: P29965 (human CD40L), Q60994 (mouse Adiponectin)

Specifications

Our Abpromise guarantee covers the use of ab168061 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Biological activity
    ab168061 binds to Human CD40.
    ab168061 induces B cells activation (as demonstrated by dose-dependent upregulation of CD86).
  • Applications

    Functional Studies

    SDS-PAGE

  • Endotoxin level
    < 0.100 Eu/µg
  • Purity
    >90% by SDS-PAGE.
    ab168061 is purified by multi-step chromatography.
  • Form
    Lyophilised
  • Additional notes

    ab168061 is a high-activity construct in which two trimeric CD40L molecules are artificially linked via the collagen domain of Adiponectin. This construct very effectively simulates the natural membrane-assisted aggregation of CD40L in vivo.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at -20ºC.

    Constituent: 99% PBS

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • Reconstitution
    Reconstitute with 100µl sterile water for a final concentration of 0.1mg/ml. After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles. Further dilutions should be made with medium containing 5% fetal calf serum.

General Info

  • Alternative names
    • CD 40L
    • CD154
    • CD40 antigen ligand
    • CD40 ligand
    • CD40 ligand, soluble form
    • CD40-L
    • CD40L
    • CD40L_HUMAN
    • CD40LG
    • gp39
    • hCD40L
    • HIGM1
    • IGM
    • IMD3
    • T B cell activating molecule
    • T BAM
    • T-cell antigen Gp39
    • TNF-related activation protein
    • TNFSF5
    • TrAP
    • Tumor necrosis factor (ligand) superfamily member 5
    • Tumor necrosis factor ligand superfamily member 5
    see all
  • Function
    Mediates B-cell proliferation in the absence of co-stimulus as well as IgE production in the presence of IL-4. Involved in immunoglobulin class switching.
    Release of soluble CD40L from platelets is partially regulated by GP IIb/IIIa, actin polymerization, and an matrix metalloproteinases (MMP) inhibitor-sensitive pathway.
  • Tissue specificity
    Specifically expressed on activated CD4+ T-lymphocytes.
  • Involvement in disease
    Defects in CD40LG are the cause of X-linked immunodeficiency with hyper-IgM type 1 (HIGM1) [MIM:308230]; also known as X-linked hyper IgM syndrome (XHIM). HIGM1 is an immunoglobulin isotype switch defect characterized by elevated concentrations of serum IgM and decreased amounts of all other isotypes. Affected males present at an early age (usually within the first year of life) recurrent bacterial and opportunistic infections, including Pneumocystis carinii pneumonia and intractable diarrhea due to cryptosporidium infection. Despite substitution treatment with intravenous immunoglobulin, the overall prognosis is rather poor, with a death rate of about 10% before adolescence.
  • Sequence similarities
    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    The soluble form derives from the membrane form by proteolytic processing.
    N-linked glycan is a mixture of high mannose and complex type. Glycan structure does not influence binding affinity to CD40.
    Not O-glycosylated.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt

Images

  • SDS-PAGE analysis of ab168061.
  • B cell lymphocyte activation by ab168061.

    Method: Peripheral Blood Mononuclear Cells (PBMCs) were incubated at 37°C, 5% CO2 for 48 hours in 48 well plates (1 x 106 cells/well). Each well contained 200µl serum free test media with serially diluted ab168061. After treatment, cells were washed and dual-stained with mouse anti-human CD19–PE and mouse anti-human CD86–APC and analyzed by flow cytometry. The data are presented as the percent of CD19 positive B cells that co-stain as CD86 positive, at each concentration of ab168061.

References

ab168061 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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