Human AFP PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human AFP 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 AFP
Protein name: Alpha-fetoprotein
Gene Full Name: alpha fetoprotein
abbreviation: AFP
Background: Alpha-fetoprotein (AFP, α-fetoprotein; also sometimes called alpha-1-fetoprotein, alpha-fetoglobulin, or alpha fetal protein) is a protein that in humans is encoded by the AFP gene. The AFP gene is located on the q arm of chromosome 4 (4q25). AFP is the most abundant plasma protein found in the human fetus. Plasma levels decrease rapidly after birth but begin decreasing prenatally starting at the end of the first trimester. Normal adult levels are usually achieved by the age of 8 to 12 months. The function of AFP in adult humans is unknown; however, in rodents it binds estradiol to prevent the transport of this hormone across the placenta to the fetus. The main function of this is to prevent the virilization of female fetuses. As human AFP does not bind estrogen, its function in humans is less clear.
ID:P02771
ID:174

alias

Afp|Alpha fetoglobulin|Alpha-fetoglobulin|Alpha fetoprotein|Alpha-fetoprotein|Alpha-1-fetoprotein|FETA|HPAFP

Origin

Nativ and recombinant human AFP

Antibodies

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

Immunogene

NSO, R19-V609

sensitivity

<10pg/ml

detection range

78pg/ml-5000pg/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)|784|1507|3564|817|1785|4516
Standard deviation|49.39|97.96|185.33|65.36|130.31|365.80
CV(%)|6.3|6.5|5.2|8|7.3|8.1

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 AFP ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for AFP has been precoated onto 96-well plates. Standards (Expression system for standard: NSO, Immunogen sequence: R19-V609) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for AFP 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 AFP amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of 5000pg/ml, 2500pg/ml, 1250pg/ml, 625pg/ml, 313pg/ml, 156pg/ml, 78pg/ml human AFP standard solutions into the pre-coated 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. We recommend that each human AFP standard solution and each sample is measured in duplicate.

tipical result

Concentration(pg/ml)|0|78|156|312|625|1250|2500|5000 O.D.|0.003|0.137|0.248|0.486|0.777|1.260|1.704|1.925

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human AFP antibody|1
ST0000-5|Lyophilized recombinant human AFP standard|5ng/tube�2
AR1107|Biotinylated anti- human AFP antibody|130µl(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