Luteinizing Hormone (LH) ELISA Kit

Details for Product No. ABIN365711, Supplier: Log in to see
Rat (Rattus)
Kits with alternative reactivity to:
Method Type
Competition ELISA
Detection Range
0.3-60 mIU/mL
Minimum Detection Limit
0.3 mIU/mL
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Supplier Product No.
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Purpose For the quantitative determination of rat luteotropic hormone (LH) concentrations in serum, plasma.
Sample Type Serum, Plasma
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This assay has high sensitivity and excellent specificity for detection of rat LH.
Cross-Reactivity (Details) Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between the target antigen and all analogues for other species. Therefore, cross reaction may still exist.
Sensitivity 0.15 mIU/mL
  • Assay plate (12 × 8 coated Microwells)
  • Standard (freeze dried)
  • Biotin-antibody (100 × concentrate)
  • HRP-avidin (100 × concentrate)
  • Biotin-antibody Diluent
  • HRP-avidin Diluent
  • Sample Diluent
  • Wash Buffer (25 × concentrate)
  • TMB Substrate
  • Stop Solution
  • Adhesive Strip (for 96 wells)
  • Instruction manual
Material not included
  • Microplate reader capable of measuring absorbance at 450nm, with the correction wavelength set at 540nm or 570nm.
  • An incubator which can provide stable incubation conditions up to 37°C ± 0.5°C.
  • Squirt bottle, manifold dispenser or automated microplate washer.
  • Absorbent paper for blotting the microtiter plate.
  • 100mL and 500mL graduated cylinders.
  • Deionized or distilled water.
  • Pipettes and pipette tips.
  • Test tubes for dilution.
Application Notes
  • The supplier is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Samples to be used within 5 days may be stored at 2-8°C, otherwise samples must be stored at -20°C (≤ 1 month) or -80°C (≤ 2 months) to avoid loss of bioactivity and contamination.
  • Grossly hemolyzed samples are not suitable for use in this assay.
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts of certain chemicals.
  • Owing to the possibility of mismatching between antigens from another resource and antibodies used in this supplier's kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by this supplier's products.
  • Influenced by factors including cell viability, cell number and cell sampling time, samples from cell culture supernatant may not be recognized by the kit.
  • Fresh samples without long time storage are recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.

Detection wavelength: 450 nm

Information on standard material:
Some standards are from natural resource, some are recombinant protein, but the recombinant protein will not expressed from Baculovirus. Generally it's CHO cells, but that depends.The formulation of auxiliary material in the standard is classified, but it dosen't contain any poisonous substance. Proclin 300 (1:3000) is used as preservative.

Information on reagents:
Most of the stop solution are 1 N H2SO4, a few is not. The formulation of wash solution is classified. None of the components contains (sodium) azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials. Some could contain BSA. For the sandwich method kits, the sample diluent, antibody diluent, enzyme diluent and standard all contain BSA.

Information on antibodies:
The provided antibodies and their host vary in different kits. Some are affinity purified, some are Protein A or G purified

Sample Volume 50 μL
Assay Time 1 - 4.5 h
Plate Pre-coated
Protocol This assay employs the competitive inhibition 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.
Sample Collection
  • Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4 °C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °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 or -80 °C. Avoid repeated freeze-thaw cycles.
  • Tissue Homogenates: Rinse 100 mg tissue with 1× PBS, homogenize in 1mL of 1× PBS and store overnight at -20 °C. After two freeze-thaw cycles to break the cell membranes, centrifuge the homogenates for 5 minutes at 5000 × g, 2-8 °C. Remove and assay the supernate immediately. Alternatively, aliquot and store samples at -20 °C or -80 °C. Centrifuge the sample again after thawing before the assay. Avoid repeated freeze-thaw cycles.
Calculation of Results

Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the target antigen concentration versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

Assay Precision Intra-assay precision (precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess precision.
Inter-assay precision (precision between assays): Three samples of known concentration were tested in twenty assays to assess precision.
  • Intra-assay: CV% less than 8%
  • Inter-assay: CV% less than 10%
Restrictions For Research Use only
Precaution of Use The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
Handling Advice
  • The kit should not be used beyond the expiration date on the kit label.
  • Do not mix or substitute reagents with those from other lots or sources.
  • If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.
  • Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time/temperature and kit age can cause variation in binding.
  • This assay is designed to eliminate interference by soluble receptors, binding proteins and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
Storage 4 °C/-20 °C
Storage Comment May be stored at 2-8°C for up to 1 month. For long term storage, please store at -20°C. Try to keep assay plate in a sealed aluminium foil bag and avoid dampness.
Expiry Date 6 months
Supplier Images
ELISA image for Luteinizing Hormone (LH) ELISA Kit (ABIN365711) Typical standard curve
Product cited in: Haron, Mohamed: "Effect of honey on the reproductive system of male rat offspring exposed to prenatal restraint stress." in: Andrologia, 2015 (PubMed).

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Sepehrimanesh, Saeb, Nazifi, Kazemipour, Jelodar, Saeb: "Impact of 900 MHz electromagnetic field exposure on main male reproductive hormone levels: a Rattus norvegicus model." in: International journal of biometeorology, Vol. 58, Issue 7, pp. 1657-63, 2014 (PubMed).

Karamikheirabad, Behzadi, Faghihi, Raoofian, Ejtemaei Mehr, Zuure, Sadeghipour: "A role for endocannabinoids in acute stress-induced suppression of the hypothalamic-pituitary-gonadal axis in male rats." in: Clinical and experimental reproductive medicine, Vol. 40, Issue 4, pp. 155-62, 2014 (PubMed).

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Uzun, Atli, Perk, Burukoglu, Ilgin: "Evaluation of the reproductive toxicity of naproxen sodium and meloxicam in male rats." in: Human & experimental toxicology, 2014 (PubMed).

López-Doval, Salgado, Pereiro, Moyano, Lafuente: "Perfluorooctane sulfonate effects on the reproductive axis in adult male rats." in: Environmental research, Vol. 134, pp. 158-68, 2014 (PubMed).

Kumar, Kaur: "Intermittent fasting dietary restriction regimen negatively influences reproduction in young rats: a study of hypothalamo-hypophysial-gonadal axis." in: PLoS ONE, Vol. 8, Issue 1, pp. e52416, 2013 (PubMed).

Kaya, Sogut, Cayli, Suren, Arici, Karaman, Erdemir: "Evaluation of effects of repeated sevoflurane exposure on rat testicular tissue and reproductive hormones." in: Inhalation toxicology, Vol. 25, Issue 4, pp. 192-8, 2013 (PubMed).

Ramezani Tehrani, Noroozzadeh, Zahediasl, Ghasemi, Piryaei, Azizi: "Prenatal testosterone exposure worsen the reproductive performance of male rat at adulthood." in: PLoS ONE, Vol. 8, Issue 8, pp. e71705, 2013 (PubMed).

Zin, Omar, Khan, Musameh, Das, Kassim: "Effects of the phytoestrogen genistein on the development of the reproductive system of Sprague Dawley rats." in: Clinics (São Paulo, Brazil), Vol. 68, Issue 2, pp. 253-62, 2013 (PubMed).

Bharti, Misro, Rai: "Clomiphene citrate potentiates the adverse effects of estrogen on rat testis and down-regulates the expression of steroidogenic enzyme genes." in: Fertility and sterility, Vol. 99, Issue 1, pp. 140-8, 2012 (PubMed).

Meena, Misro, Ghosh, Nandan: "Extended intervention time and evaluation of sperm suppression by dienogest plus testosterone undecanoate in male rat." in: Contraception, Vol. 85, Issue 1, pp. 113-21, 2011 (PubMed).

Shindel, Xin, Lin, Fandel, Huang, Banie, Breyer, Garcia, Lin, Lue: "Erectogenic and neurotrophic effects of icariin, a purified extract of horny goat weed (Epimedium spp.) in vitro and in vivo." in: The journal of sexual medicine, Vol. 7, Issue 4 Pt 1, pp. 1518-28, 2010 (PubMed).

Alsiö, Birgner, Björkblom, Isaksson, Bergström, Schiöth, Lindblom: "Impact of nandrolone decanoate on gene expression in endocrine systems related to the adverse effects of anabolic androgenic steroids." in: Basic & clinical pharmacology & toxicology, Vol. 105, Issue 5, pp. 307-14, 2009 (PubMed).