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Rat KiSS-1 ELISA Kit

SKU:
RTFI00469
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q7TSB7
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
Kiss1, Kisspeptin-1
Reactivity:
Rat
$779
Frequently bought together:

Description

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Rat KiSS-1 ELISA Kit

The Rat KiSS-1 ELISA Kit is a high-quality assay designed for the precise measurement of KiSS-1 levels in rat samples, including serum, plasma, and tissue homogenates. With its high sensitivity and specificity, this kit provides accurate and reliable results for researchers studying the KiSS-1 protein and its role in various physiological processes.KiSS-1, also known as metastin, is a neuropeptide that plays a crucial role in the regulation of reproduction, metastasis, and energy metabolism.

Dysregulation of KiSS-1 expression has been implicated in conditions such as infertility, cancer, and obesity, making it a valuable biomarker for investigating these complex diseases and potential therapeutic interventions.The Rat KiSS-1 ELISA Kit is a valuable tool for researchers looking to study the impact of KiSS-1 on physiological and pathological processes in rat models, providing a comprehensive understanding of its function and potential therapeutic implications.

Product Name:Rat Kiss1 (Metastasis-suppressor KiSS-1) ELISA Kit
Product Code:RTFI00469
Size:96 Assays
Target:Rat Kiss1
Alias:Kiss1, Kisspeptin-1
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat Kiss1 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat Kiss1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10194
EDTA plasma(n=5)87-10596
UFH plasma(n=5)92-10196
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat Kiss1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-98%85-103%85-96%
EDTA plasma(n=5)83-101%82-101%86-98%
UFH plasma(n=5)81-86%81-100%82-94%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:Q7TSB7
UniProt Protein Function:KiSS-1: Metastasis suppressor protein in malignant melanomas and in some breast cancers. May regulate events downstream of cell- matrix adhesion, perhaps involving cytoskeletal reorganization. Generates a C-terminally amidated peptide, metastin which functions as the endogenous ligand of the G-protein coupled receptor GPR54. Activation of the receptor inhibits cell proliferation and cell migration, key characteristics of tumor metastasis. Kp-10 is a decapeptide derived from the primary translation product, isolated in conditioned medium of first trimester trophoblast. Kp-10, but not other kisspeptins, increased intracellular Ca(2+) levels in isolated first trimester trophoblasts. Kp-10 is a paracrine/endocrine regulator in fine- tuning trophoblast invasion generated by the trophoblast itself. The receptor is also essential for normal gonadotropin-released hormone physiology and for puberty. The hypothalamic KiSS1/GPR54 system is a pivotal factor in central regulation of the gonadotropic axis at puberty and in adulthood. Belongs to the KISS1 family.
UniProt Protein Details:

Protein type:Secreted; Secreted, signal peptide; Cell adhesion

Cellular Component: apical plasma membrane; cell soma; cytoplasm; cytosol; extracellular space; neuron projection

Molecular Function:G-protein-coupled receptor binding; kisspeptin receptor binding

Biological Process: elevation of cytosolic calcium ion concentration; elevation of cytosolic calcium ion concentration during G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); G-protein coupled receptor protein signaling pathway; generation of ovulation cycle rhythm; negative regulation of cell migration; negative regulation of cell proliferation; positive regulation of growth hormone secretion; positive regulation of luteinizing hormone secretion; positive regulation of MAPKKK cascade; positive regulation of stress-activated MAPK cascade; positive regulation of synaptic transmission

NCBI Summary:ligand for the G-protein-coupled receptor GPR54; may be involved in autonomic and sensory neural signaling [RGD, Feb 2006]
UniProt Code:Q7TSB7
NCBI GenInfo Identifier:32140194
NCBI Gene ID:289023
NCBI Accession:NP_859043.1
UniProt Related Accession:Q7TSB7
Molecular Weight:14,188 Da
NCBI Full Name:metastasis-suppressor KiSS-1
NCBI Synonym Full Names:KiSS-1 metastasis-suppressor
NCBI Official Symbol:Kiss1  
NCBI Official Synonym Symbols:Eseptin  
NCBI Protein Information:metastasis-suppressor KiSS-1
UniProt Protein Name:Metastasis-suppressor KiSS-1
UniProt Synonym Protein Names:Kisspeptin-1
Protein Family:Metastasis-suppressor
UniProt Gene Name:Kiss1  
UniProt Entry Name:KISS1_RAT
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.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37°C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

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 clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum 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 anti-coagulant. 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 & homogenizein 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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