Overview

  • Product name
    Human CD40L ELISA Kit
    See all CD40L kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 6 pg/ml
  • Range
    8.23 pg/ml - 6000 pg/ml
  • Recovery

    95 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 96.35 86% - 105%
    Serum 93.57 83% - 103%
    Plasma 94.63 84% - 105%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Human CD40L ELISA Kit (ab99991) is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human CD40L in serum, plasma (EDTA or heparin), and cell culture supernatants.


    This assay employs an antibody specific for Human CD40L coated on a 96-well plate. Standards and samples are pipetted into the wells and CD40L present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human CD40L antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of CD40L bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get results in 90 minutes with Human CD40 Ligand ELISA Kit (TNFSF5) (ab196268) from our SimpleStep ELISA® range.


     

  • Notes

    Optimization may be required with urine samples.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human CD40L (lyophilized) 2 vials
    CD40L Microplate (12 x 8 well strips) 1 unit
    Recombinant Human CD40L Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas
  • 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
  • Alternative names
    • CD154
    • CD40 antigen ligand
    • CD40 ligand
    • CD40-L
    • CD40L
    • CD40L_HUMAN
    • CD40LG
    • gp39
    • hCD40L
    • HIGM1
    • IGM
    • IMD3
    • soluble form
    • T B cell activating molecule
    • T BAM
    • T cell antigen Gp39
    • T-cell antigen Gp39
    • TNF related activation protein
    • TNF-related activation protein
    • TNFSF5
    • TrAP
    • Tumor necrosis factor (ligand) superfamily member 5
    • Tumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome)
    • Tumor necrosis factor ligand superfamily member 5
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab99991 in the following tested applications.

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

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Representative Standard Curve using ab99991.

  • Representative Standard Curve using ab99991.

Protocols

References

ab99991 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us. This kit does recognize the extracellular region of CD40L (aa 47 – 261). Thus, it is used for the detection of soluble CD40L. The exact cross-reactivity of membrane-bound CD40L has not been tested. Please let me know if you have further questions.

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Question
Answer

Merci de votre patience.
Le laboratoire m'avait envoyé les données qu'ils ont obtenu avec ce lot. Veuillez les trouves dans le document attaché. Les résultats sont mieux que ceux que vous avez obtenue avec le kit que nous vous avions envoyé. Si vous voulez, je peux donc vous envoyer un nouveau kit comme remplacement gratuit. C'est à vous de décider. Comme discuté dans le dernier émail, je ne conseille pas d'utiliser le sérum pure, mais au plus concentré à 1/3. Pour avoir une sensitivité maximale du kit, nous vous recommandons aussi de faire très attention au pipetage et faire assez de lavages. Aussi, nous pouvons recommander de centrifuger les échantillons avant l'utilisation pour enlever des agrégats éventuelles.
Comme de toute façon les valeurs de CD40L sont très basses dans les échantillons, vous pourriez aussi penser à concentrer les échantillons (sous conditions non-dénaturants, comme avec des spin colums) et de faire l'incubation des échantillons sur la nuit à 4°C.
J'espère que ces informations vous sont utiles. Veuillez me confirmer comment vous souhaitez procéder ' si vous souhaitez un kit gratuit de remplacement ou si vous souhaitez plutôt un avoir ou remboursement.
Veuillez ne pas hésiter à nous appeler pour discuter ce problème et les options. Je me réjouis de vos nouvelles.

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Question
Answer

Thank you for contacting us. We unfortunately can’t provide aspecific starting dilution factorat whichthere will be no interference from other plasma proteins in the assay.We recommend trying a series of dilutions until a linear response is measured and determined empirically. I have also had chatwithmy colleague from lab; they have suggestedthat because plasmasamples have differences based on source and method of isolation soevery sample dilution factor should be determined separately. I am sorry we can’t be of more help in this case; the dilution factor has to be determined by the end user. Ihope this information will be helpful. Should you have any other question please do not hesitate to contactus.

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Answer

Thank you for your reply. As I mentioned in my previous email, we do not have a recommended minimum dilution for this kit because the amount of dilution necessary will depend on your samples.  Our recommended starting dilution for further optimization is 2 fold.  I am sorry I cannot provide a more specific recommendation, please let me know if you need any additional information. 

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Answer

Thank you for contacting us.  Matrix effects can sometimes occur when certain factors in the plasma such as auto-antibodies, binding proteins, albumin, etc. can interfere with the assay if too much is present in your samples.  The result is that the sample may read falsely high or low, or may give a non-linear dilution response.   I hope this helps, please let me know if you need any additional information.     

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Answer

Merci de nous avoir contactés. J'ai reçu les informations suivantes concernant les kits ELISA CD40L et PF4 : ab99991 doit être utilisé avec des échantillons de plasma traités EDTA ou héparine. L'utilisation d'autres anticoagulants n'est pas recommandée. ab100628 peut être utilisé avec des échantillons de plasma traités EDTA, héparine ou citrate. Concernant les conditions de stockage, le laboratoire n'a pas d'information spécifique pour CD40 et PF4 dans le plasma. En règle générale, les protéines contenues dans le sérum ou le plasma sont stables à -80°C pour une durée d'environ un an. Préparation recommandée des échantillons : - Plasma : Collecter le sang un tube d'EDTA ou d'héparine. Centrifuger à 3000g pendant 10 minutes. Aliquoter et stocker à -80°C - Sérum : Collecter le sang sans additif. Maintenir à température ambiante pendant 20 minutes. Centrifuger à 3000g pendant 10 minutes. Aliquoter et stocker à -80°C. ------------------------ ab99991 should only be used with EDTA-plasma or heparin-plasma. Use of other anti-coagulants is not recommended. ab100628 can be used with EDA-plasma, heparin-plasma or citrate-plasma. FYI, unless otherwise stated in the Introduction Section of the manual, if the kit is compatible with plasma, it will be compatible with heparin, EDTA, and citrate anti-coagulants. As far as plasma sample storage, I’m not aware of a stability study specifically for CD40 ligand and Platelet Factor 4 in plasma, but in general most protein should be stable in serum or plasma for at least a year if stored at -80ºC. For Plasma : Collect blood into EDTA or Sodium heparin tube Spin 10 minutes at 3000g Aliquot into small tubes and store at –80°C until use. For serum: Collect blood into tube without additive Keep at room temperature for 20 minutes Spin 10 minutes at 3000g Aliquot into small tubes and store at –80°C until use.

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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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