The Rat 8-OHdG (8-Hydroxydeoxyguanosine) ELISA Kit is specifically designed for the precise measurement of 8-hydroxydeoxyguanosine levels in rat samples, including serum, plasma, and cell culture supernatants. With its exceptional sensitivity and specificity, this kit ensures accurate and consistent results, making it a valuable tool for various research applications.8-Hydroxydeoxyguanosine is a key biomarker of oxidative stress and DNA damage, with implications in various diseases such as cancer, neurodegenerative disorders, and cardiovascular diseases.
By measuring 8-OHdG levels, researchers can gain insights into the extent of oxidative damage and assess the effectiveness of potential therapies targeting oxidative stress.Overall, the Rat 8-OHdG ELISA Kit offers researchers a reliable and robust solution for studying oxidative damage and its role in disease development and progression. With its user-friendly protocol and high performance, this kit is a valuable asset for advancing research in the field of oxidative stress and DNA damage.
Product Name:
Rat 8-OHdG (8-Hydroxydeoxyguanosine) ELISA Kit
Product Code:
RTFI01271
Size:
96T
Alias:
8-OHdG
Detection method:
Competitive ELISA, Coated with Antigen
Application:
8-OHdG ELISA Kit allows for the in vitro quantitative determination of 8-OHdG concentrations in serum, plasma, tissue homogenates and other biological fluids.
Sensitivity:
< 0.281ng/ml
Range:
0.468-30ng/ml
Storage:
2-8°C for 6 months
Note:
For Research Use Only
Recovery:
Matrices listed below were spiked with certain level of 8-OHdG and the recovery rates were calculated by comparing the measured value to the expected amount of 8-OHdG in samples.
Matrix
Recovery range(%)
Average(%)
serum(n=5)
86-105
92
EDTA plasma(n=5)
90-103
97
heparin plasma(n=5)
88-102
96
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 8-OHdG and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample
1:2
1:4
1:8
serum(n=5)
90-101%
83-95%
84-100%
EDTA plasma(n=5)
89-98%
82-97%
84-92%
heparin plasma(n=5)
87-105%
82-99%
85-100%
CV(%):
Intra-Assay: CV<8% Inter-Assay: CV<10%
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 clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum 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 anticoagulant. 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 & homogenize in 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.
Marius Fuchs et al.
Oxidative stress in rat brain during experimental status epilepticus: effect of antioxidants
