Name: Protein A (PA) ELISA Kit (NEGEP0890)
Brand: Shanghai BlueGene Biotech CO., LTD.
Price: 100 USD

BlueGene Biotech Protein A (PA) ELISA Kit (NEGEP0890)

Immunology

NEGES0890 Protein A (PA) ELISA Kit

This kit is for the quantitative Measurement of Protein A Residues in Protein Purification Process, and End-Product (purified fermentation broth, cell culture supernatant, etc.)

Products

Specifications of Protein A (PA) ELISA Kit

Product Information
Cat. NO.	NEGEP0890
Product Name	Protein A (PA) ELISA Kit
Species	General
Product Size	96 Tests
Concentration	31.25-2000pg/ml
Sensitivity	LOD: 3.9pg/mL LOQ: 31.25pg/mL
Principal	Sandwich ELISA
Sample Volume	100 ul
Sample Type	ELISA Kit for the quantitative Measurement of Protein A Residues in Protein Purification Process, and End-Product (purified fermentation broth, cell culture supernatant, etc.)
Assay Time	3 hours
Platform	Microplate Reader
Conjugate	HRP
Detection Method	Colorimetric
Storage	2-8°C

Kit Components
Reagents	Specification	Quantity
Pre-Coated Microplate (Detachable)	96 wells	1 plate (Keep Sealed)
Standard (Stock Solution) -1ug/ml	20ul	1 vial
HRP-conjugated antibody(400×)	150ul	1 vial
Sample Pretreatment Solution	5ml	1 vial
TMB Substrates	10ml	1 vial (Avoid Light)
Stop Solution	10ml	1 vial
Wash Solution (100×)	10ml	1 vial
Diluent Buffer(10x)	10ml	1 vial
Detection Antibody Diluent (1×)	10mL	2 vials
Plate Sealer		4 pieces
Instruction Manual		1

Principle of the Assay

This ELISA kit is applied to the double antibody sandwich Enzyme Linked Immunosorbent Assay to detect the concentration of protein A in samples. Before starting the ELISA experiment, mix the samples and serially diluted standards with denaturing solution, and incubate at room temperature to completely dissociate Protein A from the antibodies. After pretreatment, add the detection antibody to the microplate wells pre-coated with chicken anti-Protein A antibody. Then add the denatured standards and samples. Incubate at room temperature to allow the specific binding of antigen-antibody, capturing Protein A from the samples and standards onto the microplate. After incubation, a sandwich complex forms with the coating antibody, Protein A, and detection antibody. Wash the plate to remove unbound substances. Then a TMB substrate solution is added to the wells to incubate. TMB substrate solution turns blue after the oxidation of HRP, and finally turns to yellow immediately after adding the stop solution. Color of TMB substrate positively correlated with total protein A bound in the initial steps. The color is measured by spectrophotometrically with wavelength of 450nm. The concentration of protein A in samples is then determined by comparing the O.D. of the samples to the standard curve.

Quality Control on Protein A (PA) ELISA Kit

Coefficient of Variance	Intra Variation: 7-8%
	Inter Variation: 6-9%
Recovery Rate	In the first experiments, the matrix effect on experimental results should be confirmed through spike recovery. It is recommended to add one volume of S1(2000pg/mL) to every three volumes of the sample. Calculate the recovery rate using the formula: (actual concentration after spiking - concentration before spiking) / theoretical concentration (500pg/mL). A recovery rate of 80-120% is considered acceptable.Matrix sources that interfere with sample detection include extreme pH (pH < 5 or pH > 8.5), high salt concentrations, and detergents, which can lead to low recovery rates. High concentrations of antibody products may sometimes interfere with assay results. It is recommended to pre-dilute such high-concentration samples prior to testing.