Rat GM-CSF PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of rat GM-CSF in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).

spezie

Rat

Cross Reaction

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

detection target

Rat GM-CSF
Protein name: Granulocyte-macrophage colony-stimulating factor
Gene Full Name: colony stimulating factor 2 (granulocyte-macrophage)
abbreviation: CSF2
Background: Granulocyte-macrophage colony-stimulating factor(GM-CSF) is also symbolized CSF2. Human GM-CSF is a glycoprotein that is essential for the in vitro proliferation and differentiation of precursor cells into mature granulocytes and macrophages. The human cDNA clones contain a single open-reading frame encoding a protein of 144 amino acids with a predicted molecular mass of 16,293 daltons and show 69% nucleotide homology and 54% amino acid homology to mouse GM-CSF. The gene for human GM-CSF appears to exist as a single-copy gene. Human GM-CSF is a 22,000-dalton glycoprotein that stimulates the growth of myeloid progenitor cells and acts directly on mature neutrophils. The GM-CSF gene is localized by somatic cell hybrid analysis and in situ hybridization to human chromosome region 5q21-5q32, which is involved in interstitial deletions in the 5q- syndrome and acute myelogenous leukemia. A complementary DNA for the T lymphocyte-derived lymphokine, GM-CSF has been cloned, and recombinant GM-CSF protein has been expressed in yeast and purified to homogeneity. This purified human recombinant GM-CSF stimulates peripheral blood monocytes in vitro to become cytotoxic for the malignant melanoma cell line A375.
ID:P48750
ID:116630

alias

Colony stimulating factor| Colony stimulating factor 2 (granulocyte macrophage)| CSF| CSF 2| CSF2| GMCSF| Granulocyte macrophage colony stimulating factor| MGC131935| MGC138897| Molgramostin| Sargramostim

Origin

Nativ and recombinant rat GM-CSF

Antibodies

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

Immunogene

E.coli, A1-K127

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)|110|379|646|125|458|637
Standard deviation|4.29|16.3|29.07|8.5|31.6|45.2
CV(%)|3.9|4.3|4.5|6.8|6.9|7.1

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 rat GM-CSF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for GM-CSF has been precoated onto 96-well plates. Standards(E.coli, A1-K127) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for GM-CSF 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 rat GM-CSF 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 rat GM-CSF 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 rat 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 rat GM-CSF 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.|0.030|0.085|0.138|0.229|0.423|0.806|1.391|2.271

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- rat GM-CSF antibody|1
ST0000-10|Lyophilized recombinant rat GM-CSF standard|10ng/tubex2
AR1107|Biotinylated anti- rat GM-CSF antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(dilution 1:100)
AR1106-1|Sample diluent buffer|30 ml
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