Human CD28 PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human CD28 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 CD28
Protein name: T-cell-specific surface glycoprotein CD28
Gene Full Name: CD28 molecule
abbreviation: CD28
Background: CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co- stimulatory signals required for T cell activation and survival. T cell stimulation through CD28 in addition to the T-cell receptor (TCR) can provide a potent signal for the production of various interleukins (IL-6 in particular). The CD28 gene is mapped to 2q33-q34. CD28 and CTLA-4, together with their ligands, B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T and B cell responses. CD28 and CTLA-4 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily.
ID:P10747
ID:940

alias

CD 28 | CD28 | CD28 antigen | CD28 molecule | TP44 | P10747

Origin

Nativ and recombinant human CD28

Antibodies

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

Immunogene

NSO, N19-P152

sensitivity

<12pg/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(pg/ml)|482|1431|2547|498|1554|3041
Standard deviation|21.69|72.98|147.73|35.86|99.46|206.79
CV(%)|4.5|5.1|5.8|7.2|6.4|6.8

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 CD28 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD28 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: N19-P152) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD28 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 CD28 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 CD28 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 CD28 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.|0.023|0.114|0.235|0.399|0.776|1.301|1.790|2.021

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human CD28 antibody|1
ST0000-10|Lyophilized recombinant human CD28 standard|10ng/tube�2
AR1107|Biotinylated anti- human CD28 antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(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