QuicKey Human IgM(Immunoglobulin M) ELISA Kit

SKU: E-TSEL-H0010

$ 331⁹⁵

QuicKey Human IgM(Immunoglobulin M) ELISA Kit

Detection Range	7.80-500 ng/mL
Sensitivity	1.02 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	7.80-500 ng/mL
Sensitivity	1.02 ng/mL
Sample type &Sample volume	serum, plasma, urine, saliva; 50μL
Specificity	This kit recognizes Human IgM in samples. No significant or interference between Human IgM and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human IgM concentrations in serum, plasma, urine, saliva.Please consult technical support for the applicability if other biological fluids need to be tested.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IgM. Samples (or Standards) and biotinylated detection antibody specific for Human IgM are added to the micro ELISA plate wells. Human IgM would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IgM, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human IgM. You can calculate the concentration of Human IgM in the samples by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	2-8℃ (Protect from light)
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IgM were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IgM were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	20.40	57.60	229.31	21.61	62.32	248.67
Standard deviation	1.21	2.63	12.59	1.11	3.00	12.86
CV (%)	5.93	4.57	5.49	5.14	4.81	5.17

Recovery

The recovery of Human IgM spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	93-101	98
EDTA plasma(n=8)	97-106	100
Urine(n=8)	80-112	104

Target Information

Background

Immunoglobulin M (IgM) is a class of immunoglobulins found in blood and lymph fluid that are the first type of antibody made in response to an infection [1]. IgM antibodies make up about 5 percent to 10 percent of all the antibodies in the body. IgM is principally concerned with the primary antibody response, appearing soon after initial invasion by an antigen and capable of destroying the antigen when it is first introduced. Its normal mean of serum concentration is 120 mg/dl. IgM is the first immunoglobulin expressed in the human fetus (around 20 weeks) and phylogenetically the earliest antibody to develop [2]. IgM antibodies appear early in the course of an infection and usually reappear, to a lesser extent, after further exposure. IgM antibodies do not pass across the human placenta (only isotype IgG). The biological properties of IgM make it useful in the diagnosis of infectious diseases. Demonstrating IgM antibodies in a patient's serum indicates recent infection, o
1. Gupta S, Pahwa R, O'Reilly R, et al. Ontogeny of lymphocyte subpopulations in human fetal liver. [J]. Proceedings of the National Academy of Sciences of the United States of America, 1976, 73(3):919-922.
2. Van F R, Schuit H R, Hijmans W. The immunological development of the human fetus. [J]. Journal of Experimental Medicine, 1965, 122(6):1173.

Research Area

Cancer, Immunology

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results