Overview

  • Product name
    Human CD40 ELISA Kit
    See all CD40 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 5 5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 5%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    4.9 pg/ml
  • Range
    31.25 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 100 98% - 104%
    Saliva 91 88% - 95%
    Milk 100 98% - 104%
    Urine 98 93% - 100%
    Serum 98 92% - 102%
    Cell culture extracts 97 90% - 101%
    Heparin Plasma 103 101% - 105%
    EDTA Plasma 100 97% - 103%
    Citrate Plasma 91 87% - 94%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    CD40 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CD40 protein in human serum, plasma, milk, saliva, urine, cell culture supernatants, and cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:
    Samples diluted in Sample Diluent NS MDD = 4.9 pg/mL
    Samples diluted in 1X Cell Extraction Buffer PTR MDD = 6.7 pg/mL

  • Notes

    CD40 is single-pass type I cell membrane glycoprotein that belongs to the TNF receptor superfamily. CD40 participates in normal immune system development and function. Alternative splicing generates CD40 isoform II that lacks the transmembrane domain and is secreted. Besides alternative splicing, soluble CD40 can be also generated by proteolytic cleavage of transmembrane CD40. CD40 is expressed on variety of cells including B cells, dendritic cells, monocytes, macrophages, T cells, neutrophils, platelets, fibroblasts, smooth muscle cells, epithelial cells, endothelial cells, neurons, and in many hematopoietic and epithelial cancer cells. CD40 is a receptor for TNFSF5/CD40LG. Both CD40 and CD40LG are upregulated on many of these cell types during inflammation and/or activation. CD40 homotrimerization transduces TRAF6- and MAP3K8-mediated signals that activate ERK in macrophages and B cells, leading to induction of immunoglobulin secretion.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human CD40 Capture Antibody 1 x 600µl
    10X Human CD40 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BI 1 x 6ml
    Human CD40 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas
  • Function
    Receptor for TNFSF5/CD40LG.
  • Tissue specificity
    B-cells and in primary carcinomas.
  • Involvement in disease
    Defects in CD40 are the cause of hyper-IgM immunodeficiency syndrome type 3 (HIGM3) [MIM:606843]; also known as hyper-IgM syndrome 3. HIGM3 is an autosomal recessive disorder which includes an inability of B cells to undergo isotype switching, one of the final differentiation steps in the humoral immune system, an inability to mount an antibody-specific immune response, and a lack of germinal center formation.
  • Sequence similarities
    Contains 4 TNFR-Cys repeats.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Alternative names
    • AI326936
    • B cell associated molecule CD40
    • B cell surface antigen CD40
    • B cell-associated molecule
    • B-cell surface antigen CD40
    • Bp50
    • CD 40
    • CD40
    • CD40 antigen
    • CD40 antigen (TNF receptor superfamily member 5)
    • CD40 molecule
    • CD40 molecule, TNF receptor superfamily member 5
    • CD40 protein
    • CD40 type II isoform
    • CD40L receptor
    • CDw40
    • GP39
    • HIGM1
    • IGM
    • IMD3
    • MGC9013
    • Nerve growth factor receptor related B lymphocyte activation molecule
    • OTTHUMP00000031699
    • OTTHUMP00000031700
    • p50
    • T-BAM
    • TBAM
    • TNF receptor superfamily member 5
    • TNFRSF5
    • TNR5_HUMAN
    • TRAP
    • Tumor necrosis factor receptor superfamily , member 5
    • Tumor necrosis factor receptor superfamily member 5
    • Tumor necrosis factor receptor superfamily member 5 precursor
    • Tumor necrosis factor receptor superfamily, member 5, isoform CRA_a
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab210970 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

  • ELISA Protocol Summary
  • Standard curve comparison between mouse CD40 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity with shorter protocol time.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of CD40 were measured in duplicates, interpolated from the CD40 standard curves and corrected for sample dilution. Undiluted samples are as follows:  urine 20%, MDA-MB-231 supernatant 100%, milk 12.5%, and saliva 12.5%.  The interpolated dilution factor corrected CD40 concentration in neat (100%) samples are plotted (mean +/- SD, n=2).  The mean CD40 concentration was determined to be 9170 pg/mL in urine, 226 pg/mL in MDA-MB-231 supernatant, 1238 pg/mL in milk, and 637 pg/mL in saliva.

  • The concentrations of CD40 were measured in duplicate and interpolated from the CD40 standard curve and corrected for sample dilution.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean CD40 concentration was determined to be 1,199 pg/mL in MDA-MB-231 cell extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean CD40 concentration was determined to be 877 pg/mL with a range of 568 – 1,562 pg/mL

Protocols

References

ab210970 has not yet been referenced specifically in any publications.

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