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MAK322

Collagen Assay Kit

sufficient for 100 fluorometric tests

Synonym(s):

Collagen Quantification Kit

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About This Item

NACRES:
NA.84
UNSPSC Code:
12161503
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usage

sufficient for 100 fluorometric tests

input

cell
serum
plasma
solution(s) (collagen)

application(s)

cosmetics
food and beverages

detection method

fluorometric

relevant disease(s)

rheumatological diseases; dermatological diseases; gastrointestinal diseases; pulmonary disorders; cancer

storage temp.

−20°C

General description

Collagen is the key structural protein of connective tissue and the most abundant protein in mammals. It occurs in many different types and forms with Types I -V being the most common. Aside from the crucial role it plays in the body, it has numerous medical applications such as its use in reconstructive surgery including bone and skin grafts. It is also commonly used in cosmetics due to its anti-aging and skin healing properties.

Application

Suitable for biological samples (e.g. plasma, serum, cells) and cosmetic samples (e.g. collagen solution)
The Collagen Assay Kit may be used for:
  • Bone Disorder Research
  • Dermatological Research
  • Wound Healing and Tissue Repair Research


Biochem/physiol Actions

Assay methods available for quantifying collagen currently range from needing extensive hydrolysis procedures with acids and bases to using expensive antibodies and complicated protocols. The collagen assay kit delivers a very simple, safe (non-radioactive), and sensitive method to quantify collagen in samples. In the first step of this procedure, collagen in the sample is enzymatically digested into peptides. Subsequently, the N-terminal glycine containing peptides react with the dye reagent to form a fluorescent complex. The fluorescence intensity of this product, measured at λex/em = 375/465 nm, is directly proportional to collagen concentration in the sample.

Features and Benefits

Enhanced Detection Range: Detect Collagen levels accurately across a wide range, from 2 µg/mL to 50 µg/mL, using only 20 μL of sample in a 96-well plate setup, ensuring highly sensitive analysis with minimal sample requirement.

Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.

Other Notes

For additional information on our range of Biochemicals, please complete this form.

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Collagen Assay Kit sufficient for 100 fluorometric tests

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MAK322

Collagen Assay Kit

Soluble Collagen Quantification Assay Kit

Sigma-Aldrich

CS0006

Soluble Collagen Quantification Assay Kit

Succinate Assay Kit sufficient for 100 colorimetric or fluorometric tests

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3D Collagen Culture Kit 3D cell culture kit includes: Human Collagen, Mediums (RPMI, M199 and DMEM), PBS with Phenol Red and Neutralization Solution

Sigma-Aldrich

ECM675

3D Collagen Culture Kit

detection method

fluorometric

detection method

-

detection method

colorimetric, fluorometric

detection method

-

usage

sufficient for 100 fluorometric tests

usage

sufficient for 200 fluorometric tests

usage

sufficient for 100 colorimetric or fluorometric tests

usage

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

2-8°C

input

cell
serum
plasma
solution(s) (collagen)

input

-

input

beverage(s)
food(s)
biological sample(s)

input

-

application(s)

cosmetics
food and beverages

application(s)

-

application(s)

-

application(s)

-

relevant disease(s)

rheumatological diseases; dermatological diseases; gastrointestinal diseases; pulmonary disorders; cancer

relevant disease(s)

-

relevant disease(s)

cardiovascular diseases; immunological diseases

relevant disease(s)

-

pictograms

Health hazard
GHS08

signalword

Danger

Hazard Classifications

Repr. 1B - Resp. Sens. 1

Storage Class

6.1D - Non-combustible acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects

wgk

WGK 2

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Questions

1–10 of 10 Questions  

  1. What is the appropriate cell lysis method for this assay?

    1 answer

    1. The recommended cell lysis method for this assay is sonication to avoid the use of detergents. However, a detergent such as Triton X-100 is expected to be compatible with the assay. An example of a cell lysis protocol for 0.4-2.0 x 10^5 HeLa cells is as follows:

      Harvest cells by centrifugation at 2,000 x g for 5 minutes at 4°C or by trypsinization for adherent cells.
      Homogenize or sonicate cells in PBS for ten minutes.
      Centrifuge at 10,000 x g for 5 minutes at 4°C.
      Remove the supernatant for the assay.
      If a lysis buffer is preferred, avoid Tris-based buffers as they may interfere with the assay. Additionally, the following substances should be avoided as they may inhibit the kit's collagenase: EDTA, EGTA, cysteine, histidine, DTT, 2-mercaptoethanol, o-phenanthroline, Hg2+, Pb2+, Cd2+, and Cu2+. The assay is designed to detect type 1 collagen. Although type 3 collagen has not been explicitly tested, it is expected to be detected as well due to structural similarities.

      Helpful?

  2. Can I use this kit to stain collagen and perform imaging under a microscope?

    1 answer

    1. This product has been optimized and is intended for use in a fluorescent plate reader.

      Helpful?

  3. How can I determine the shelf life / expiration / retest date of this product?

    1 answer

    1. If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
      For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
      For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
      For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/418/501/product-dating-information-06-25-mk.pdf

      Helpful?

  4. Does this assay detect gelatin?

    1 answer

    1. The is kit utilizes collagenase which will cleave both collagen and gelatin (denatured collagen). However, this is will not differentiate between collagen and gelatin.

      Helpful?

  5. How is shipping temperature determined? And how is it related to the product storage temperature?

    1 answer

    1. Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf

      Helpful?

  6. Does this kit detect total collagen?

    1 answer

    1. Yes, this kit quantifies total collagen but does not distinguish between specific types.

      Helpful?

  7. Does this kit detect total collagen?

    1 answer

    1. Yes, this kit quantifies total collagen, but does not distinguish between specific types.

      Helpful?

  8. Would the MAK322 Collagen Assay Kit be suitable for use with whole cell lysates containing RIPA buffer?

    1 answer

    1. No tests have been conducted on the effect of RIPA on this assay. It is believe that RIPA-treated cell lysates should be compatible with the kit, as long as they do not contain chelation agents such as EDTA. However, they discourage the use of chelators because the collagenase used in the assay requires calcium as a cofactor.

      Helpful?

  9. Will the assay work if the samples to be dosed are either or both pH’s, and do they have to be neutralized? If so, to what pH (from both extremes)? Also, Does the collagen standard also have to be blanked?

    1 answer

    1. 1. The samples should be neutralized before assaying. The assay buffer is around pH 7.5, so it is recommended that samples be neutralized to around pH 7 to 8.
      2. The collagen standard does not have to be blanked. During calculation, standard 4 (only water) is subtracted from the other standards as a blank.

      Helpful?

  10. Is this kit compatible with cell lysates in detergent buffers (ex. RIPA buffer)?

    1 answer

    1. The compatibility of RIPA in this assay has not been evaluated. However, no interference is anticipated. The use of any additional chelating agents, such as EDTA, are not recommended and will negatively impact results.

      Helpful?

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