GFP PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of GFP in cell lysates and tissue homogenates.

spezie

Pan-species

Cross Reaction

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

detection target

GFP
Protein name: Green fluorescent protein
Background: The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. In cell and molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms it has been used to make biosensors, and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, injection with a viral vector, or cell transformation. To date, the GFP gene has been introduced and expressed in many Bacteria, Yeast and other Fungi, fish (such as zebrafish), plant, fly, and mammalian cells, including human.
ID:P42212

alias

GFP|Green fluorescent protein|yfp

Origin

Nativ and recombinant GFP

Antibodies

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

Immunogene

E.coli, S2-K238

sensitivity

<10pg/ml

detection range

15.6pg/ml-1000pg/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)|96|351|668|115|427|722
Standard deviation|4.61|24.22|41.42|6.44|30.32|53.43
CV(%)|4.8|6.9|6.2|5.6|7.1|7.4

TMB(tetramethylbenzidine) incubation time

25-30

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 GFP ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for GFP has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: S2-K238) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for GFP 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 GFP amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.3pg/ml, 15.6pg/ml GFP 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 cell lysates or tissue homogenates to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. We recommend that each GFP standard solution and each sample is measured in duplicate.

tipical result

Concentration(pg/ml)|0|15.6|31.2|62.5|125|250|500|1000 O.D.|0.055|0.137|0.182|0.295|0.496|0.897|1.563|2.225

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- GFP antibody|1
ST0000-10|Lyophilized recombinant GFP standard|10ng/tube�2
AR1107|Biotinylated anti- GFP 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