Horse AMH (Anti-Mullerian Hormone) ELISA Kit (AEFI01061)

Product Name:	Horse AMH (Anti-Mullerian Hormone) ELISA Kit
Alias:	Muellerian-inhibiting factor ELISA Kit, Anti-Muellerian hormone ELISA Kit, AMH ELISA Kit, Muellerian-inhibiting substance ELISA Kit, MIS ELISA Kit, AMH ELISA Kit, MIF ELISA Kit
SKU:	AEFI01061
Size:	96T
Species:	Horse
Sample Type:	Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method:	Competitive ELISA, Coated with Antigen
Detection Wavelength:	OD450
Reaction Duration:	2 hours
Range:	0.156-10ng/ml
Sensitivity:	0.094ng/ml
Storage:	2-8°C(Sealed), Don't cryopreserve.
Specificity:	Specifically binds with AMH, no obvious cross reaction with other analogues.

This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with AMH. AMH in the sample or standard competes with a fixed amount of biotin-labeled AMH for sites on a pre-coated antibody specific to AMH. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), biotin specifically binds with streptavidin to form immune complexes. Wash off unbound conjugate. Add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of AMH in the sample. The concentration of the target substance is inversely proportional to the OD450 value.

Component	Size (96T)	Storage
ELISA Microplate (Dismountable)	8×12	Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C
Lyophilized Standard	2 vials	Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C
Biotin-labeled Antibody (Concentrated, 100X)	60 ul	2-8°C (Avoid Direct Light)
HRP-Streptavidin Conjugate (SABC, 100X)	120 ul
TMB Substrate	10 ml
Sample Dilution Buffer	20 ml	2-8°C
Antibody Dilution Buffer	10 ml
SABC Dilution Buffer	10 ml
Stop Solution	10 ml
Wash Buffer (Concentrated, 25X)	30 ml
Plate Sealer	5 pieces
Product Description	1 copy

Other materials and equipment required:

Microplate reader (wavelength: 450nm)
37°C incubator (CO2 incubator for cell culture is not recommended.)
Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips (Calibration is required before use.)
Sterile tubes and Eppendorf tubes with disposable tips
Absorbent paper and loading slot
Deionized or distilled water

Recovery:

Add a certain amount of AMH into the sample. Calculate the recovery by comparing the measured value with the expected amount of AMH in the sample.

Sample Type	Recovery Range(%)	Average(%)
Serum(n=5)	85-103	92
EDTA Plasma(n=5)	93-103	98
Heparin Plasma(n=5)	86-105	95

Linearity:

Dilute the sample with a certain amount of AMH at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
Serum(n=5)	86-104%	85-100%	85-98%
EDTA Plasma(n=5)	85-104%	86-100%	87-100%
Heparin Plasma(n=5)	86-98%	88-97%	85-97%

Stability:

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5)	37°C for 1 month	2-8°C for 6 months
Average(%)	80	95-100

Precision(%):

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/ml)	0.31	1.24	5.2	0.35	1.16	5.16
Standard deviation	0.02	0.06	0.22	0.02	0.06	0.24
CV(%)	5.22	4.69	4.23	5.29	5.26	4.58

Step 1:	Wash the plate twice before adding the standard and sample.
Step 2:	Add 50ul standard or sample into each well. Immediately add 50ul biotin-labeled antibody working solution into each well (The tip can't be reused after touching the liquid.), gently shake the plate for 1 min, seal closely and statically incubate for 45 minutes at 37°C.
Washing:	Wash the plate three times and immerse for 1min each time.
Step 3:	Add 100ul HRP-Streptavidin Conjugate (SABC) working solution into each well, seal the plate and statically incubate for 30 minutes at 37°C.
Washing:	Wash the plate five times and immerse for 1min each time.
Step 4:	Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 5:	Add 50ul stop solution. Read at 450nm immediately and calculate.