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Rat AQP4 / Aquaporin-4 ELISA Kit

SKU:
RTFI00211
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P47863
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
AQP-4, Aquaporin-4, AQP4, HMIWC2, MIWC, WCH4
Reactivity:
Rat
Research Area:
Signal Transduction
€649
Frequently bought together:

Description

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Rat AQP4/Aquaporin-4 ELISA Kit

The Rat AQP4 (Aquaporin-4) ELISA Kit is specifically designed for the accurate measurement of Aquaporin-4 levels in rat samples, including serum, plasma, and cell culture supernatants. This kit boasts high sensitivity and specificity, ensuring precise and reproducible results for various research applications.Aquaporin-4 is a key protein involved in water transportation in the central nervous system and plays a crucial role in maintaining brain water homeostasis.

Dysregulation of AQP4 has been linked to various neurological disorders such as neuromyelitis optica and brain edema, making it a valuable biomarker for understanding these conditions and developing potential treatments.Overall, the Rat AQP4 ELISA Kit is an essential tool for researchers studying the role of Aquaporin-4 in neurological diseases and water balance regulation in the brain.

Product Name:Rat AQP-4 (Aquaporin-4) ELISA Kit
Product Code:RTFI00211
Size:96 Assays
Target:Rat AQP-4
Alias:AQP-4, Aquaporin-4, AQP4, HMIWC2, MIWC, WCH4
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 AQP-4 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat AQP-4 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10595
EDTA plasma(n=5)85-10394
UFH plasma(n=5)85-10495
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat AQP-4 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)91-97%87-104%87-101%
EDTA plasma(n=5)83-89%86-98%88-100%
UFH plasma(n=5)84-94%83-94%80-95%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P47863
UniProt Protein Function:AQP4: a member of the aquaporin family of intrinsic membrane proteins that function as water-selective channels in the plasma membranes of many cells. The predominant aquaporin found in brain. Two alternatively spliced transcript variants have been described
UniProt Protein Details:

Protein type:Channel, misc.; Membrane protein, multi-pass; Transporter, aquaporin family; Transporter; Membrane protein, integral

Cellular Component: basal plasma membrane; basolateral plasma membrane; cell projection membrane; cytoplasm; external side of plasma membrane; integral to membrane; integral to plasma membrane; intercellular junction; membrane; plasma membrane; pore complex; protein complex; sarcolemma; T-tubule

Molecular Function:glycerol channel activity; porin activity; transporter activity; water channel activity

Biological Process: carbon dioxide transport; female pregnancy; glycerol transport; hyperosmotic salinity response; multicellular organismal water homeostasis; negative regulation of interleukin-1 beta production; negative regulation of interleukin-6 production; protein homooligomerization; response to glucocorticoid stimulus; response to radiation; sensory perception of sound; transmembrane transport; transport; water homeostasis; water transport

NCBI Summary:This gene encodes a member of the aquaporin family of intrinsic membrane proteins that function as water-selective channels in the plasma membranes of many cells. This protein is the predominant aquaporin found in brain and has an important role in brain water homeostasis. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. A recent study provided evidence for translational readthrough in this gene and expression of an additional C-terminally extended isoform via the use of an alternative in-frame translation termination codon. [provided by RefSeq, Dec 2015]
UniProt Code:P47863
NCBI GenInfo Identifier:395759219
NCBI Gene ID:25293
NCBI Accession:NP_001257487.1
UniProt Related Accession:P47863
Molecular Weight:28,768 Da
NCBI Full Name:aquaporin-4 isoform 3
NCBI Synonym Full Names:aquaporin 4
NCBI Official Symbol:Aqp4  
NCBI Official Synonym Symbols:Miwc; WCH4; AQP-4  
NCBI Protein Information:aquaporin-4
UniProt Protein Name:Aquaporin-4
UniProt Synonym Protein Names:Mercurial-insensitive water channel; MIWC; WCH4
Protein Family:Aquaporin
UniProt Gene Name:Aqp4  
UniProt Entry Name:AQP4_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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