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Mouse SHBG / Sex Hormone Binding Globulin ELISA Kit

SKU:
MOFI01108
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P97497
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Competitive
Synonyms:
SHBG, ABP, androgen binding protein, SBP, sex hormone-binding globulin, Sex steroid-binding protein, TeBG, TEBG, Testis-specific androgen-binding protein, testosterone-binding beta-globulin, Testosterone-estradiol-binding globulin
Reactivity:
Mouse
€649
Frequently bought together:

Description

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Mouse SHBG/Sex Hormone Binding Globulin ELISA Kit

The Mouse SHBG (Sex Hormone Binding Globulin) ELISA Kit is specifically designed for the accurate measurement of SHBG levels in mouse serum, plasma, and cell culture supernatants. This kit provides high sensitivity and specificity, ensuring precise and consistent results, making it perfect for various research applications.Sex Hormone Binding Globulin (SHBG) is a key protein that binds to sex hormones, regulating their availability and activity in the body.

Changes in SHBG levels have been associated with various health conditions such as hormonal imbalances, metabolic disorders, and reproductive issues, making it a valuable biomarker for studying these conditions and developing potential treatments.Overall, the Mouse SHBG ELISA Kit is an essential tool for researchers looking to investigate the role of SHBG in different physiological processes and diseases in mouse models.

Product Name:Mouse SHBG / Sex Hormone Binding Globulin ELISA Kit
Product Code:MOFI01108
Size:96 Assays
Alias:SHBG, ABP, androgen binding protein, SBP, sex hormone-binding globulin, Sex steroid-binding protein, TeBG, TEBG, Testis-specific androgen-binding protein, testosterone-binding beta-globulin, Testosterone-estradiol-binding globulin
Detection Method:Competitive ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse SHBG concentrations in serum plasma and other biological fluids.
Sensitivity:18.75pg/ml
Range:31.25-2000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery: Matrices listed below were spiked with certain level of Mouse SHBG and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse SHBG in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)92-10398
EDTA plasma(n=5)92-10298
UFH plasma(n=5)85-10291
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse SHBG and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)94-105%89-100%86-103%
EDTA plasma(n=5)83-101%88-100%89-101%
UFH plasma(n=5)80-96%85-95%90-100%
Intra Assay:CV <8%
Inter Assay:CV <10%
ComponentQuantityStorage
ELISA Microplate(Dismountable)8-12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)60ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP97497
UniProt Protein Function:Functions as an androgen transport protein, but may also be involved in receptor mediated processes. Each dimer binds one molecule of steroid. Specific for 5-alpha-dihydrotestosterone, testosterone, and 17-beta-estradiol. Regulates the plasma metabolic clearance rate of steroid hormones by controlling their plasma concentration.
UniProt Code:P97497
NCBI GenInfo Identifier:6755506
NCBI Gene ID:20415
NCBI Accession:NP_035497.1
UniProt Related Accession:P97497
Molecular Weight:44 kDa
NCBI Full Name:sex hormone-binding globulin
NCBI Synonym Full Names:sex hormone binding globulin
NCBI Official Symbol:Shbg  
NCBI Official Synonym Symbols:ABP  
NCBI Protein Information:sex hormone-binding globulin
UniProt Protein Name:Sex hormone-binding globulin
UniProt Synonym Protein Names:Sex steroid-binding protein; SBP; Testis-specific androgen-binding protein; ABP
Protein Family:Sex hormone-binding globulin
UniProt Gene Name:Shbg  

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2. Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blank well is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody working solution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thorough mixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming).
3. Wash: Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 350µL) using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC working solution to each well. Cover with a new Plate sealer. Incubate for 30 minutes at 37°C.
5.Wash: Repeat the aspiration/wash process for five times.
6. TMB Substrate: Add 90µL of TMB Substrate to each well. Cover with a new Plate sealer. Incubate for about 10-20 minutes at 37°C. Protect from light. The reaction time can be shortened or extended according to the actual color change, but not more than 30minutes. When apparent gradient appeared in standard wells, you can terminate the reaction.
7. Stop: Add 50µL of Stop Solution to each well. Color turn to yellow immediately. The adding order of stop solution should be as the same as the substrate solution.
8. OD Measurement: Determine the optical density (OD Value) of each well at once, using a microplate reader set to 450 nm. You should open the microplate reader ahead, preheat the instrument, and set the testing parameters.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum:

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

Plasma:

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 - g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid:

Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant:

Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.

Cell lysates:

Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C.

Tissue homogenates:

The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.

Tissue lysates:

Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.

Breast Milk:

Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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