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Rat TRY4 ELISA Kit

SKU:
RTFI00447
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P12788
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
TRY4, Pretrypsinogen IV, Trypsin IV,
Reactivity:
Rat
Research Area:
Cell Biology
€649
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Description

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Rat TRY4 ELISA Kit

The Rat TRY4 ELISA Kit from Assay Genie is a powerful tool for researchers looking to accurately measure levels of trypsinogen 4 (TRY4) in rat samples. This ELISA kit offers high sensitivity and specificity, ensuring precise and reliable results for a variety of research applications.TRY4 is an important enzyme involved in the digestive process, specifically in the activation of trypsinogen into trypsin. Abnormal levels of TRY4 have been linked to various gastrointestinal disorders, making it a valuable biomarker for studying these conditions and developing potential therapies.

With the Rat TRY4 ELISA Kit, researchers can confidently analyze rat serum, plasma, and cell culture supernatants to gain valuable insights into TRY4 levels and its implications in gastrointestinal health. Trust Assay Genie's Rat TRY4 ELISA Kit for accurate and reproducible results in your research endeavors.

Product Name:Rat TRY4 (Trypsin-4) ELISA Kit
Product Code:RTFI00447
Size:96 Assays
Target:Rat TRY4
Alias:TRY4, Pretrypsinogen IV, Trypsin IV
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 TRY4 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat TRY4 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)93-10397
EDTA plasma(n=5)87-10094
UFH plasma(n=5)93-10398
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat TRY4 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-97%89-103%89-98%
EDTA plasma(n=5)82-100%83-98%89-97%
UFH plasma(n=5)82-95%82-96%86-99%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P12788
NCBI Summary:member of the trypsin family of serine proteases [RGD, Feb 2006]
UniProt Code:P12788
NCBI GenInfo Identifier:27545370
NCBI Gene ID:286960
NCBI Accession:NP_775423.1
UniProt Related Accession:P12788
Molecular Weight:26.1 kDa
NCBI Full Name:trypsin-4
NCBI Synonym Full Names:preprotrypsinogen IV
NCBI Official Symbol:LOC286960  
NCBI Official Synonym Symbols:Try4  
NCBI Protein Information:trypsin-4
UniProt Protein Name:Trypsin-4
UniProt Synonym Protein Names:Pretrypsinogen IV; Trypsin IV
Protein Family:Transcriptional regulator of yeast form adherence
UniProt Gene Name:Try4  
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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