High Throughput Superoxide Dismutase Assay Kit

R&D Systems | Catalog # 7501-500-K

Determines relative SOD activity by measuring the percent inhibition of the rate of formation of WST-1 formazan.
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Key Product Details

Features

Analyzes Superoxide Dismutase (SOD) activity in cell extracts using a 96-well plate.

Species

Multi-Species

Product Summary for High Throughput Superoxide Dismutase Assay Kit

Why Use the HT Superoxide Dismutase Assay Kit?
In the HT Superoxide Dismutase Assay Kit, superoxide radical (O2 –) ions, generated from the conversion of xanthine to uric acid and H2 O2 by xanthine oxidase (XOD), convert WST-1 to WST-1 Formazan. WST-1 Formazan absorbs light at 450 nm. SODs reduce superoxide ion concentrations and thereby lower the rate of WST-1 formazan formation. The extent of reduction in the appearance of WST-1 Formazan is a measure of SOD activity present in your experimental sample. Superoxide dismutases (SODs) catalyze the dismutation of the superoxide radical (O2 –) into hydrogen peroxide (H2 O2). The HT Superoxide Dismutase Assay Kit is free of interference by other catalytic activities and is ideal for assaying SOD in mammalian tissue and cell lysates in a 96 well format. Unlike some other assay kits for SOD, this system is minimally disturbed by trace metals. Sufficient reagents are provided for 480 tests. The assay is performed in as little as 6 minutes and relative SOD activity of the experimental sample is determined as the percent inhibition of the rate of formation of WST-1 Formazan. This assay is also suitable for the assay of isozymes SOD1 (cytosolic Cu/Zn-SOD), SOD2 (mitochondrial Mn-SOD), and SOD3 (extracellular Cu/Zn-SOD).
Other Resources
7501-500-k_calculation_worksheet

Kit Contents for High Throughput Superoxide Dismutase Assay Kit

  • SOD Standard
  • Xanthine Oxidase (XOD)
  • 10X SOD Buffer
  • 10X Xanthine Solution
  • 20% Triton X-100
  • WST-1 Reagent
  • 96-Well Plate

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended on the product label.

Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: SOD (Superoxide Dismutase)

Superoxide Dismutases, originally identified as Indophenoloxidases (IPOs), are enzymes that catalyze the conversion of naturally-occuring but harmful superoxide radicals into molecular oxygen and hydrogen peroxide. Three mammalian isozymes of SOD have been identified and are functionally related but have very modest sequence homology. SODs are typically soluble secreted or cytosolic proteins, but are also found in the mitochondria and extracellular matrix.

Any of three metals, manganese, iron, or copper, may be used in the active site of SOD and are indicative of cellular localization.MnSOD (SOD2) is found in the mitochondria of both prokaryotes and ukaryotes, whereas the cytosol of eukaryotes and prokaryotic organisms contains Cu/ZnSOD (SOD1) and FeSOD, respectively.

There have been several mutations identified in the Cu/ZnSOD (SOD1) gene in amyotrophic lateral sclerosis (ALS) patients, possibly suggesting a role for free radicals in this disease process. The mechanism of motor neuron degeneration that occurs in ALS, however, is unclear at this time. Several hypotheses have been explored for the role of mutant SOD1 and convincing evidence for the involvement of apoptosis has been presented, as well.

Long Name

Superoxide Dismutase

Alternate Names

SOD

Additional SOD (Superoxide Dismutase) Products

Product Documents for High Throughput Superoxide Dismutase Assay Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Citations for High Throughput Superoxide Dismutase Assay Kit

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FAQs for High Throughput Superoxide Dismutase Assay Kit

Showing  1 - 5 of 6 FAQs Showing All

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

  • Q: Can frozen tissues be used in this assay?

    A: We have not tested frozen tissue in this assay. The protocol is based on isolating SOD from fresh tissue. We recommend isolating the cytosolic extracts prior to freezing the samples. Fresh tissue is recommended because single cell suspension from  tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. 

  • Q: Can this kit be used to measure saliva samples?

    A: This kit may be used to measure SOD activity in saliva samples within the detection range of the assay.

  • Q: Can this kit be used to measure serum or plasma samples?

    A: This kit may be used to measure SOD activity in serum or plasma samples within the detection range of the assay.

  • Q: Is it necessary to determine protein concentration of the cleared cell lysate, as instructed in the protocol  for the HT Superoxide Dismutase Assay (catalog # 7501-500-K)?

    A: Determination of total protein concentration of the sample whose SOD activity being measured, allows to measure the specific activity of the enzyme in that sample. See page 11 of the product insert for example calculation.

  • Q: What is the Lower Limit of Detection (LLD) of this kit?

    A: An exact LLD has not been established for this kit. However, the lowest amount that is recommended for the standard curve is 0.1 Unit of SOD. The standard curve is based on a purified enzyme.

  • Q: Which form of SOD does this kit detect?

    A: This kit detects all SOD forms.

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