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Rat Deoxyribonuclease-1 / DNASE1 ELISA Kit

SKU:
RTFI00314
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P21704
Sensitivity:
0.234ng/ml
Range:
0.391-25ng/ml
ELISA Type:
Sandwich
Synonyms:
Dnase1, DNase-I
Reactivity:
Rat
Research Area:
Cell Death
€649
Frequently bought together:

Description

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Rat Deoxyribonuclease-1/DNASE1 ELISA Kit

The Rat Deoxyribonuclease 1 (DNase1) ELISA Kit is a cutting-edge tool designed for the precise measurement of DNase1 levels in rat serum, plasma, and cell culture supernatants. This kit boasts exceptional sensitivity and specificity, ensuring the delivery of accurate and reproducible results for a variety of research applications.DNase1 is a crucial enzyme involved in DNA degradation and maintenance of genomic integrity. Its dysregulation has been linked to various diseases such as autoimmune disorders, inflammation, and tissue damage.

This ELISA kit serves as a valuable tool for researchers studying these conditions and developing potential therapeutic interventions.With its user-friendly protocol and reliable performance, the Rat DNase1 ELISA Kit is an indispensable resource for scientists seeking to elucidate the intricate role of DNase1 in health and disease in rat models. Elevate your research with this advanced kit and unravel the mysteries of DNase1 biology.

Product Name:Rat Dnase1 (Deoxyribonuclease-1) ELISA Kit
Product Code:RTFI00314
Size:96 Assays
Target:Rat Dnase1
Alias:Dnase1, DNase-I
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.234ng/ml
Range:0.391-25ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat Dnase1 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat Dnase1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)95-10398
EDTA plasma(n=5)87-9491
UFH plasma(n=5)85-10593
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat Dnase1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-101%85-102%87-102%
EDTA plasma(n=5)85-101%84-98%84-98%
UFH plasma(n=5)80-91%84-100%80-82%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P21704
UniProt Protein Function:Among other functions, seems to be involved in cell death by apoptosis. Binds specifically to G-actin and blocks actin polymerization.
NCBI Summary:glycoprotein that cleaves DNA endonucleolytically to yield 5(alpha)-phosphodinucleotide and oligonucleotide end products; facilitates DNA hydrolysis in ailementary duct and other digestive tissues [RGD, Feb 2006]
UniProt Code:P21704
NCBI GenInfo Identifier:6978769
NCBI Gene ID:25633
NCBI Accession:NP_037229.1
UniProt Secondary Accession:P21704,Q5FVU6,
UniProt Related Accession:P21704
Molecular Weight:32,064 Da
NCBI Full Name:deoxyribonuclease-1
NCBI Synonym Full Names:deoxyribonuclease I
NCBI Official Symbol:Dnase1  
NCBI Protein Information:deoxyribonuclease-1
UniProt Protein Name:Deoxyribonuclease-1
UniProt Synonym Protein Names:Deoxyribonuclease I; DNase I
Protein Family:Deoxyribonuclease
UniProt Gene Name:Dnase1  
UniProt Entry Name:DNAS1_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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