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Home / ELISA Kit / PharmaTest ELISA Kits

Cetuximab (Erbitux) Pharmacokinetic ELISA

SKU : BHE18300003

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Mfr.No.	EL-1611-031
Detection Range	50ng/ml - 1.56ng/ml
Reactivity	Human, Mouse, Rat
Assay Duration	2.5 hours
Assay Type	Direct, Sandwich
Description	Cetuximab (Erbitux ) is a chimeric IgG1 monoclonal antibody that binds the extra-cellular domain of the epidermal growth factor receptor (EGFR). It is a 152-kDa molecule composed of four polypeptide chains: two identical heavy chains and two identical light chains, consisting of 449 and 214 amino acids, respectively, bound by covalent and non-covalent bonds. The bond with EGFR is characterized by a higher affinity than either endogenous ligand, as epidermal growth factor (EGF), or transforming growth factor alpha. This binding inhibits activation of the receptor tyrosine kinase and the associated downstream signaling that includes the mitogenactivated protein kinase, phosphoinositide 3-kinase/ Akt and the Janus kinases/ signal transducers and activator of transcription (Stat) pathways. Furthermore Cetuximab induces antibody-mediated receptor dimerization, internalization and degradation leading to receptor down-regulation. In addition, it exhibits antibody-dependent cellular cytotoxicity that could contribute to its antitumor effect. Reagent preparation Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation Dilute wash buffer concentrate with deionized water 1/10 before use (for example add 20mL concentrate to 180mL deionized water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11ul concentrate to 11ml of assay buffer). Mix well. The following is an example calibrator curve. Results calculation 1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor. Sample preparation Dilute calibrators and test samples 1/100 with assay buffer (for example add 5uL of prepared calibrator or sample to 495uL of assay buffer). Mix well. Do not store diluted samples.
Detection Method	Peroxidase / OD450
Kit Components	Coated microtiter plate, 96 wells Calibrator diluent. - 1.8ml Calibrator 12ul 10X wash buffer - 25ml Assay buffer - 50ml 1000X detection reagent - 17ul TMB - 12ml TMB stop solution - 12ml Plate sealers - 3
Limit Of Detection	1.5ng/ml
Components	Coated microtiter plate, 96 wells Calibrator diluent. - 1.8ml Calibrator 12ul 10X wash buffer - 25ml Assay buffer - 50ml 1000X detection reagent - 17ul TMB - 12ml TMB stop solution - 12ml Plate sealers - 3
Precision	Intra-assay coefficient of variation (CV) <10%. Inter-assay CV <10%.
Notes	This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20°C for up to 1 year.
Usage Notes	Quantification of Cetuximab in biological matrices
Protocol	The Cetuximab ELISA kit is designed to measure free Cetuximab with high specificity and sensitivity . This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Cetuximab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Cetuximab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Cetuximab present in test samples and the concentration is calculated from the standard series.

Gene ID	1956

Applications	ELISA

Storage	-20C, 1 year

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SKU	Supplier No.	Size	Your price	Quantity
BHE18300003	EL-1611-031	1X96 wells	$809.99		Add to Cart

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