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Rat Luteinizing hormone (LH) ELISA Kit

Citations(2)Uniprot : N/A
  • Cat.No.: AE25909RA

  • Reactivity: Rat (Rattus norvegicus)

To Purchase AE25909RA

Size:
48T 96T
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Reagent Preparation







Results demonstration







Assay Procedure Summary







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Product Details

Species Reactivity Rat (Rattus norvegicus)
UniProt N/A
Abbreviation LH
Alternative Names N/A
Range Request Information
Sensitivity Request Information
Sample Type Serum, Plasma, Other biological fluids
Detection Method Sandwich
Analysis Method Quantitive
Assay Duration 1-4.5h
Sample Volume 1-200 μL
Detection Wavelengt 450 nm

Test principle

This assay employs the competitive enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with goat-anti-rabbit antibody. Standards or samples are added to the appropriate microtiter plate wells with an antibody specific for LH and Horseradish Peroxidase (HRP) conjugated LH. The competitive inhibition reaction is launched between with HRP labeled LH and unlabeled LH with the antibody. A substrate solution is added to the wells and the color develops in opposite to the amount of LH in the sample. The color development is stopped and the intensity of the color is measured.

 

Product Overview

Luteinizing hormone (LH) is an important hormone of the reproductive system, which has found application in diagnosis and therapeutic medicine. It plays a vital role in the development and functioning of the reproductive system. Determination of LH concentration is important for detection of dysfunction of the pituitary variance axis, diagnosis of reproductive disorders, monitoring of antifertility programmes and in therapeutic preparations.
LH levels are normally low during childhood and, in women, high after menopause. As LH is secreted as pulses, it is necessary to follow its concentration over a sufficient period of time to get a proper information about its blood level.During the reproductive years typical levels are between 1-20 IU/L. Physiologic high LH levels are seen during the LH surge (v.s.); typically they last 48 hours.

 

Components


Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 10 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

 

Specificity

This assay has high sensitivity and excellent specificity for detection of Rat LH. No significant cross-reactivity or interference between Rat LH and analogues was observed.
 

Recovery

Matrices listed below were spiked with certain level of recombinant Rat LH and the recovery rates were calculated by comparing the measured value to the expected amount of Rat LH in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

 

Precision

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat LH and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

 

Stability

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
 

Sample collection and storage

Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
 

Kits storage instructions

Store at 2-8°C. Please refer to Instruction Manual.
 
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