Human sCD40L PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human soluble CD40L in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).

spezie

Human

Cross Reaction

There is no detectable cross-reactivity with other relevant proteins.

detection target

Human CD40L
Protein name: CD40 ligand
Gene Full Name: CD40 ligand
abbreviation: CD40LG
Background: CD40 ligand(CD40L) is a type II membrane protein of 261 amino acids on activated T cells that induces B cell proliferation and immunoglobulin secretion. It has homology with tumour necrosis factor-alpha and -beta, and has important functions in B-cell activation and differentiation. Human CD40L with 5 exons, is mapped to the proximal region of the mouse X chromosome on Xq26.3-27.1, and can be detected on T cells but is absent from B cells and monocytes. Since CD40L is expressed on platelets and released from them on activation, its predictive value as a marker for clinical outcome and the therapeutic effect of inhibition of glycoprotein IIb/IIIa receptor in patients with acute coronary syndromes was investigated. The soluble CD40L may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells.CD40L plays a central role in the pathophysiology of acute coronary syndromes, and has a role in the pathogenesis of coronary artery lesions.
ID:P29965
ID:959

alias

CD154|CD40 antigen ligand|CD40 ligand|CD40-L|CD40L_HUMAN|CD40LG|gp39|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

Origin

Nativ and recombinant human CD40L

Antibodies

a monoclonal antibody from mouse|a biotinylated polyclonal antibody from goat

Immunogene

E.coli, E108-L261

sensitivity

<15pg/ml

detection range

62.5pg/ml-4000pg/ml

pack size

1x96 Wells

Reproducibility

|Intra-Assay Precision|Inter-Assay Precision
Sample|1|2|3|1|2|3
n|16|16|16|24|24|24
Mean(ng/ml)|0.44|1.26|2.38|0.42|1.41|2.46
Standard deviation|0.026|0.064|0.145|0.029|0.086|0.182
CV(%)|5.9|5.1|6.1|6.8|6.1|7.4

TMB(tetramethylbenzidine) incubation time

15-20

storage and Stability

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Principle of the Elisa

Boster′s human CD40L ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD40L has been precoated onto 96-well plates. Standards(E.coli, E108-L261) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD40L is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human CD40L amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 4000pg/ml, 2000pg/ml,1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml human CD40L standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each human CD40L standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|62.5|125|250|500|1000|2000|4000 O.D.|0.084|0.148|0.194|0.337|0.518|1.035|1.572|2.111

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human sCD40L antibody|1
ST0000-10|Lyophilized recombinant human sCD40L standard|10ng/tubex2
AR1107|Biotinylated anti- human sCD40L antibody|130ul(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex(ABC)|130ul(dilution 1:100)
AR1106-1|Sample diluent buffer|30ml
AR1106-2|Antibody diluent buffer|12ml
AR1106-3|ABC diluent buffer|12ml
AR1104|TMB color developing agent|10ml
AR1105|TMB stop solution|10ml
-|Adhesive cover|4