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Rabbit Super Oxide Dismutase (SOD) ELISA Kit


Species Reactivity

RaRabbit (Oryctolagus cuniculus)

UniProt

N/A

Abbreviation

SOD

Alternative Names

ALS; ALS1; IPOA; SOD1; homodimer; Cu/Zn superoxide dismutase|SOD; soluble|indophenoloxidase A|superoxide dismutase; cystolic

Range

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Sensitivity

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Sample Type

Serum, Plasma, Other biological fluids

Detection Method

Competitive ELISA

Analysis Method

Quantitive

Assay Duration

1-4.5h

Sample Volume

1-200 μL

Detection Wavelengt

450 nm

Cat

AE64513RB

Size

48T 96T

Price

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Availability

3-5 working days

Manual

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Test principle:
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to SOD. Standards or samples are added to the appropriate microtiter plate wells with biotin-conjugated SOD. A competitive inhibition reaction is launched between PO (Standards or samples) and biotin-conjugated SOD with the pre-coated antibody specific for SOD. The more amount of SOD in samples, the less antibody bound by biotin-conjugated SOD. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Substrate solution is added to the wells and the color develops in opposite to the amount of SOD in the sample. The color development is stopped and the intensity of the color is measured.
Product Overview:
SOD outcompetes damaging reactions of superoxide, thus protecting the cell from superoxide toxicity. In biological systems, this means its main reactions are with itself (dismutation) or with another biological radical such as nitric oxide (NO) or a metal. SOD is used in cosmetic products to reduce free radical damage to skin, for example to reduce fibrosis following radiation for breast cancer. Studies of this kind must be regarded as tentative, however, as there were not adequate controls in the study including a lack of randomization, double-blinding, or placebo. SOD has proved to be highly effective in treatment of colonic inflammation in experimental colitis. Treatment with SOD decreases reactive oxygen species generation and oxidative stress and, thus, inhibits endothelial activation and indicate that modulation of factors that govern adhesion molecule expression and leukocyte-endothelial interactions.
Components:

Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 12 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

Specificity:
This assay has high sensitivity and excellent specificity for detection of Rabbit SOD1. No significant cross-reactivity or interference between Rabbit SOD1 and analogues was observed.
Recovery:
Matrices listed below were spiked with certain level of recombinant Rabbit SOD1 and the recovery rates were calculated by comparing the measured value to the expected amount of Rabbit SOD1 in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

Precision:
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rabbit SOD1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

Stability:
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
Sample collection and storage:
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Kits storage instructions:
Store at 2-8°C. Please refer to Instruction Manual.