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Rat KiSS-1 receptor (Kiss1r) ELISA Kit (RTEB1390)

SKU:
RTEB1390
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q924U1
Range:
3.12-200 ng/mL
ELISA Type:
Sandwich
Reactivity:
Rat
€649
Frequently bought together:

Description

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Rat KiSS-1 receptor (Kiss1r) ELISA Kit

The Rat Kiss-1 Receptor (KISS1R) ELISA Kit is a reliable tool for the precise measurement of KISS1R levels in rat samples such as serum, plasma, and tissue lysates. This kit is known for its high sensitivity and specificity, guaranteeing accurate and consistent results for various research applications.KISS1R, also known as GPR54, is a key receptor involved in the regulation of the reproductive system, specifically in the control of puberty and fertility. Dysregulation of KISS1R has been linked to conditions such as infertility, delayed puberty, and reproductive disorders, making it a crucial biomarker for understanding these conditions and developing potential therapeutic interventions.

Overall, the Rat Kiss-1 Receptor (KISS1R) ELISA Kit is an essential tool for researchers studying the role of KISS1R in reproductive physiology and its implications in various reproductive disorders. With its reliable performance and accurate results, this kit is a valuable asset for advancing research in this field.

Product Name:Rat KiSS-1 receptor (Kiss1r) ELISA Kit
SKU:RTEB1390
Size:96T
Target:Rat KiSS-1 receptor (Kiss1r)
Synonyms:G-protein coupled receptor 54, G-protein coupled receptor OT7T175, Kisspeptins receptor, Metastin receptor, rOT7T175, KiSS-1R, Gpr54
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:3.12-200ng/mL
Sensitivity:1.67ng/mL
Intra CV:5.2%
Inter CV:8.1%
Linearity:
Sample1:21:41:81:16
Serum(N=5)99-111%87-97%115-124%103-112%
EDTA Plasma(N=5)93-105%102-115%105-118%98-107%
Heparin Plasma(N=5)85-95%97-106%93-102%97-107%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9488-100
Plasma9690-102
Function:Receptor for metastin, a C-terminally amidated peptide of KiSS1. KiSS1 is a metastasis suppressor protein. Activation of the receptor inhibits cell proliferation and cell migration, key characteristics of tumor metastasis. The receptor is essential for normal gonadotropin-released hormone physiology and for puberty. The hypothalamic KiSS1/KISS1R system is a pivotal factor in central regulation of the gonadotropic axis at puberty and in adulthood. Analysis of the transduction pathways activated by the receptor identifies coupling to phospholipase C and intracellular calcium release through pertussis toxin-insensitive G(q) proteins.
Uniprot:Q924U1
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat KiSS-1 receptor
Subcellular Location:Cell membrane Multi-pass membrane protein
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:KISS1R: Receptor for metastin (kisspeptin-54 or kp-54), a C- terminally amidated peptide of KiSS1. KiSS1 is a metastasis suppressor protein that suppresses metastases in malignant melanomas and in some breast carcinomas without affecting tumorigenicity. The metastasis suppressor properties may be mediated in part by cell cycle arrest and induction of apoptosis in malignant cells. The receptor is essential for normal gonadotropin-released hormone physiology and for puberty. The hypothalamic KiSS1/KISS1R system is a pivotal factor in central regulation of the gonadotropic axis at puberty and in adulthood. The receptor is also probably involved in the regulation and fine- tuning of trophoblast invasion generated by the trophoblast itself. Analysis of the transduction pathways activated by the receptor identifies coupling to phospholipase C and intracellular calcium release through pertussis toxin-insensitive G(q) proteins. Defects in KISS1R are a cause of idiopathic hypogonadotropic hypogonadism (IHH). IHH is defined as a deficiency of the pituitary secretion of follicle-stimulating hormone and luteinizing hormone, which results in the impairment of pubertal maturation and of reproductive function. Defects in KISS1R are a cause of central precocious puberty (CEPREPU). Precocious puberty is defined as the development of secondary sexual characteristics before the age of 8 years in girls and 9 years in boys. Central precocious puberty refers to a gonadotropin-dependent type which results from premature activation of the hypothalamic-pituitary-gonadal axis. Belongs to the G-protein coupled receptor 1 family.Protein type: Cell cycle regulation; GPCR, family 1; Membrane protein, integral; Membrane protein, multi-pass; Receptor, GPCRCellular Component: cell surface; integral to membrane; integral to plasma membrane; intracellular; intracellular membrane-bound organelle; membrane; plasma membraneMolecular Function: G-protein coupled receptor activity; neuropeptide binding; neuropeptide receptor activity; peptide receptor activity, G-protein coupledBiological Process: activation of MAPKK activity; arachidonic acid secretion; calcium-mediated signaling; G-protein coupled receptor protein signaling pathway; G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); negative regulation of cell migration; negative regulation of cell proliferation; neuropeptide signaling pathway; positive regulation of hormone secretion; positive regulation of stress fiber formation; positive regulation of synaptic transmission; signal transduction
UniProt Protein Details:
NCBI Summary:a G protein-coupled receptor; human homolog may bind ligand and metastasis suppressor gene KiSS-1 [RGD, Feb 2006]
UniProt Code:Q924U1
NCBI GenInfo Identifier:667667733
NCBI Gene ID:78976
NCBI Accession:NP_076482.2
UniProt Secondary Accession:Q924U1,Q9Z0T7
UniProt Related Accession:Q924U1
Molecular Weight:42,889 Da
NCBI Full Name:kiSS-1 receptor isoform 1
NCBI Synonym Full Names:KISS1 receptor
NCBI Official Symbol:Kiss1r
NCBI Official Synonym Symbols:Gpr54
NCBI Protein Information:kiSS-1 receptor
UniProt Protein Name:KiSS-1 receptor
UniProt Synonym Protein Names:G-protein coupled receptor 54; G-protein coupled receptor OT7T175; rOT7T175; Kisspeptins receptor; Metastin receptor
Protein Family:KiSS-1 receptor
UniProt Gene Name:Kiss1r
UniProt Entry Name:KISSR_RAT
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

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

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. 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 experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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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