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Rat AMELX ELISA Kit

SKU:
RTFI00383
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P63278
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
Amelx
Reactivity:
Rat
Research Area:
Signal Transduction
$779
Frequently bought together:

Description

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Rat AMELX ELISA Kit

The Rat Amelogenin (AMELX) ELISA Kit is specifically designed for the precise measurement of AMELX levels in rat serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results for various research purposes.AMELX is a key protein involved in enamel development and tooth mineralization, playing a crucial role in dental health and development.

Dysregulation of AMELX has been linked to various dental disorders, making it a valuable biomarker for studying dental conditions and potential therapeutic interventions.With its superior performance and reliability, the Rat AMELX ELISA Kit is an essential tool for researchers investigating the role of AMELX in dental health and disease. Get your kit today and unlock new insights into dental biology and pathology.

Product Name:Rat Amelx (Amelogenin, X isoform) ELISA Kit
Product Code:RTFI00383
Size:96 Assays
Target:Rat Amelx
Alias:Amelx
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 Amelx and the recovery rates were calculated by comparing the measured value to the expected amount of Rat Amelx in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-9894
EDTA plasma(n=5)85-10295
UFH plasma(n=5)87-10494
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat Amelx and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-103%87-105%90-105%
EDTA plasma(n=5)82-99%83-99%84-97%
UFH plasma(n=5)86-96%84-99%81-97%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P63278
UniProt Protein Function:AMELX: Plays a role in biomineralization. Seems to regulate the formation of crystallites during the secretory stage of tooth enamel development. Thought to play a major role in the structural organization and mineralization of developing enamel. Defects in AMELX are the cause of amelogenesis imperfecta type 1E (AI1E). A X-linked defect of dental enamel formation. Teeth have only a thin layer of enamel with normal hardness. The thinness of the enamel makes the teeth appear small. Belongs to the amelogenin family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Secreted, signal peptide; Secreted

Cellular Component: extracellular matrix; Golgi apparatus; proteinaceous extracellular matrix; cell surface; endocytic vesicle; cytoplasm; basement membrane

Molecular Function:identical protein binding; growth factor activity; hydroxyapatite binding; structural constituent of tooth enamel

Biological Process: response to drug; cell proliferation; biomineral formation; cell adhesion; response to calcium ion; response to nutrient

NCBI Summary:may play a role in mineralization of tooth enamel matrix [RGD, Feb 2006]
UniProt Code:P63278
NCBI GenInfo Identifier:402692277
NCBI Gene ID:29160
NCBI Accession:NP_001258002.1
UniProt Secondary Accession:P63278,P45559, Q62945, Q63640, Q78E56,
UniProt Related Accession:P63278
Molecular Weight:21,868 Da
NCBI Full Name:amelogenin, X isoform isoform 2
NCBI Synonym Full Names:amelogenin, X-linked
NCBI Official Symbol:Amelx  
NCBI Official Synonym Symbols:Amel  
NCBI Protein Information:amelogenin, X isoform
UniProt Protein Name:Amelogenin, X isoform
UniProt Synonym Protein Names:Leucine-rich amelogenin peptide; LRAP
Protein Family:Amelogenin
UniProt Gene Name:Amelx  
UniProt Entry Name:AMELX_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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