FSH ELISA Kit

Catalog No. ABIN6955959

Product Details FSH ELISA Kit

Target: FSH (Follicle-Stimulating Hormone (FSH))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 2.47 ng/mL - 200 ng/mL

Minimum Detection Limit: 2.47 ng/mL

Application: ELISA

Purpose: The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of FSH in rat serum, plasma.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Follicle Stimulating Hormone (FSH)

Sensitivity: 1.11 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 50 μL

Assay Time: 2 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 50μL standard or sample to each well.
   Then add 50μL prepared Detection Reagent A immediately.
   Shake and mix. Incubate 1 hour at 37 °C,
3. Aspirate and wash 3 times,
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
5. Aspirate and wash 5 times,
6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
7. Add 50μL Stop Solution. Read at 450 nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 600 ng/mL. Please firstly dilute the stock solution to 200 ng/mL and the diluted standard serves as the highest standard (200 ng/mL). Then prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 200 ng/mL, 66.67 ng/mL, 22.22 ng/mL, 7.41 ng/mL, 2.47 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
4. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
5. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
7. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of causing a deviation due to the introduced chemical substance. The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Expiry Date: 6 months

References for FSH ELISA Kit (ABIN6955959)

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Zhao, Zhou, Chen, Liu, Chu, Zhang: "Beneficial effects of Heqi san on rat model of polycystic ovary syndrome through the PI3K/AKT pathway." in: Daru : journal of Faculty of Pharmacy, Tehran University of Medical Sciences, Vol. 25, Issue 1, pp. 21, (2018) (PubMed).

Wang, Wang, Liang, He, Xia, Shen, Wang, Gao, Wang: "DHEA-induced ovarian hyperfibrosis is mediated by TGF-β signaling pathway." in: Journal of ovarian research, Vol. 11, Issue 1, pp. 6, (2018) (PubMed).

Chan, Jazwiec, Gohir, Petrik, Sloboda: "Maternal nutrient restriction impairs young adult offspring ovarian signaling resulting in reproductive dysfunction and follicle loss." in: Biology of reproduction, Vol. 98, Issue 5, pp. 664-682, (2018) (PubMed).

Wang, He, Yang: "Eicosapentaenoic Acid Improves Polycystic Ovary Syndrome in Rats via Sterol Regulatory Element-Binding Protein 1 (SREBP-1)/Toll-Like Receptor 4 (TLR4) Pathway." in: Medical science monitor : international medical journal of experimental and clinical research, Vol. 24, pp. 2091-2097, (2018) (PubMed).

Zhang, Hu, Meng, Sun, Xu, Zhang, Cui, Morina, Li, Li, Wu, Brännström, Shao, Billig: "Metformin Ameliorates Uterine Defects in a Rat Model of Polycystic Ovary Syndrome." in: EBioMedicine, Vol. 18, pp. 157-170, (2017) (PubMed).

Sewani-Rusike, Ralebona, Nkeh-Chungag: "Dose- and time-dependent effects of Garcinia kola seed extract on sexual behaviour and reproductive parameters in male Wistar rats." in: Andrologia, Vol. 48, Issue 3, pp. 300-7, (2016) (PubMed).

Ashworth, George, Hogg, Lai, Brunton: "Sex-specific prenatal stress effects on the rat reproductive axis and adrenal gland structure." in: Reproduction (Cambridge, England), Vol. 151, Issue 6, pp. 709-17, (2016) (PubMed).

Tsoulis, Chang, Moore, Chan, Gohir, Petrik, Vickers, Connor, Sloboda: "Maternal High-Fat Diet-Induced Loss of Fetal Oocytes Is Associated with Compromised Follicle Growth in Adult Rat Offspring." in: Biology of reproduction, Vol. 94, Issue 4, pp. 94, (2016) (PubMed).

Zhang, Sun, Sun, Meng, Hu, Li, Li, Wu, Brännström, Shao, Billig: "Molecular characterization of insulin resistance and glycolytic metabolism in the rat uterus." in: Scientific reports, Vol. 6, pp. 30679, (2016) (PubMed).

Pampanini, Germani, Puglianiello, Stukenborg, Reda, Savchuk, Kjartansdóttir, Cianfarani, Söder: "Impact of uteroplacental insufficiency on postnatal rat male gonad." in: The Journal of endocrinology, Vol. 232, Issue 2, pp. 247-257, (2016) (PubMed).

Tiya, Sewani-Rusike, Shauli: "Effects of treatment with Hypoxis hemerocallidea extract on sexual behaviour and reproductive parameters in male rats." in: Andrologia, Vol. 49, Issue 8, (2016) (PubMed).

Sangun, Dundar, Darici, Comlekci, Doguc, Celik: "The effects of long-term exposure to a 2450?MHz electromagnetic field on growth and pubertal development in female Wistar rats." in: Electromagnetic biology and medicine, Vol. 34, Issue 1, pp. 63-71, (2015) (PubMed).

Anand, Misro, Sharma, Prakash: "Protective effects of Eugenia jambolana extract versus N-acetyl cysteine against cisplatin-induced damage in rat testis." in: Andrologia, Vol. 47, Issue 2, pp. 194-208, (2015) (PubMed).

Mihalik, Mašlanková, Solár, Horváthová, Hubková, Almášiová, Šoltés, Švaňa, Rybárová, Hodorová: "The effect of R-(-)-deprenyl administration on reproductive parameters of rat males." in: European journal of pharmacology, Vol. 754, pp. 148-52, (2015) (PubMed).

Guldvang, Hansen, Weisser, Halling-Sørensen, Styrishave: "Simvastatin decreases steroid production in the H295R cell line and decreases steroids and FSH in female rats." in: Reproductive toxicology (Elmsford, N.Y.), Vol. 58, pp. 174-83, (2015) (PubMed).

Wang, Tian, Liang, Wang, Wang, Cheng, Yan, Chen, Qi: "Mechanistic Study on Triptorelin Action in Protecting From 5-FU-Induced Ovarian Damage in Rats." in: Oncology research, Vol. 22, Issue 5-6, pp. 283-92, (2015) (PubMed).

Yigitaslan, Ozatik, Ozatik, Erol, Sirmagul, Baseskioglu: "Effects of tadalafil on hemorrhagic cystitis and testicular dysfunction induced by cyclophosphamide in rats." in: Urologia internationalis, Vol. 93, Issue 1, pp. 55-62, (2014) (PubMed).

Shahabi, Jorsaraei, Moghadamnia, Zabihi, Aghajanpour, Mousavi Kani, Pourbagher, Hosseini, Esmaili, Yoonesi, Zarghami, Alinezhad: "Central effects of camphor on GnRH and sexual hormones in male rat." in: International journal of molecular and cellular medicine, Vol. 1, Issue 4, pp. 191-6, (2014) (PubMed).

Bharti, Misro, Rai: "Quercetin supplementation restores testicular function and augments germ cell survival in the estrogenized rats." in: Molecular and cellular endocrinology, Vol. 383, Issue 1-2, pp. 10-20, (2014) (PubMed).

Target Details for FSH

Target: FSH (Follicle-Stimulating Hormone (FSH))

Alternative Name: Follicle Stimulating Hormone (FSH) (FSH Products)

Synonyms: FSH ELISA Kit, Fshbeta ELISA Kit, follicle stimulating hormone beta ELISA Kit, Plasma FSH concentration ELISA Kit, Fshb ELISA Kit, FSH ELISA Kit

Target Type: Hormone

Pathways: Peptide Hormone Metabolism, Chromatin Binding