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

BlueGene Biotech Protein A (PA) ELISA Kit (Non-Native Protein) (NEGEP0890)

Immunology

NEGES0890 Protein A (PA) ELISA Kit (Non-Native Protein)

The Protein A ELISA kit is applicable in the field of biotechnology and biomedicine, it is for quantitative measurement of Protein A residues in the purification process of biopharmaceutical production, for examples like purified fermentation broth and cell culture supernatant, etc. This ELISA kit is for research use only, it should not be used in clinical diagnostic procedures.

Products

Specifications of Protein A (PA) ELISA Kit (Non-Native Protein)

Product Information
Cat. NO.	NEGEP0890
Product Name	Protein A (PA) ELISA Kit (Non-Native Protein)
Species	General
Product Size	96 Tests
Concentration	0.1875-6ng/ml
Sensitivity	47pg/ml
Principal	Sandwich ELISA
Sample Volume	100 ul
Sample Type	ELISA Kit for the quantitative Measurement of Protein A (Non-Native Protein) 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) -2ug/ml	20ul	1 vial
Standard S1	6ng/ml	1 vial
Standard S2	3ng/ml	1 vial
Standard S3	1.5ng/ml	1 vial
Standard S4	0.75ng/ml	1 vial
Standard S5	0.375ng/ml	1 vial
Standard S6	0.1875ng/ml	1 vial
HRP-conjugated antibody(400×)	50ul	1 vial
Sample Pretreatment Solution	10ml	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
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 (Non-Native Protein)

Coefficient of Variance	Intra Variation: 7-8%
	Inter Variation: 6-9%
Recovery	In the first experiments, the matrix effect on experimental results should be confirmed through spike recovery. It is recommended to add one volume of standard solution (6 ng/mL) to every three volumes of the sample. Calculate the recovery rate using the formula: (actual concentration after spiking - concentration before spiking) / theoretical concentration (3 ng/mL). A recovery rate of 80-120% is considered acceptable.
Specificity/Cross-reactivity	Capture antibody and detection antibody respectively identify different epitopes of the antigen, which maximizes the specificity of the reaction.