Intended Use For the selective separation and enumeration of enterococci in food and water. Principle and Interpretation Tryptone and peptone are the sources of nitrogen and essential grow……
Detail
Introduction
Intended Use
For the selective separation and enumeration of enterococci in food and water.
Principle and Interpretation
Tryptone and peptone are the sources of nitrogen and essential growth factors. Yeast extract acts as well nitrogenous compounds and additionally the vitamin B12 complex. Sodium azide acts largely inhibits the growth of gram-negative bacteria while sparing enterococci, staphylococci and streptococci. Ox bile inhibits most gram positives but not enterococci. Enterococci hydrolyse esculin to esculetin and dextrose, which reacts with ferric citrate producing a brownish black precipitate around the colonies. Tolerance to bile and the ability to hydrolyze esculin is the traditional and reliable test for the identification of enterococci. (4). Sodium chloride maintains the osmotic balance of the medium and Agar is the solidifying agent.
Formulation
Ingredients
/liter
Tryptone
17.0g
Ox bile
10.0g
Yeast extract
5.0g
Sodium chloride
5.0g
Peptone
3.0g
aesculin
1.0g
Ferric ammonium citrate
0.5g
Sodium azide
0.15g
Agar
15.0g
pH 7.1±0.1 at 25°C
Preparation
Weigh 56.6g of dry powder of this product, add 1 L of distilled water or deionized water, stir, heat and boil until completely dissolved, and sterilize at 121℃ for 15 min.
Quality Control
Cultural characteristics observed after incubation at 35-37°C for 20-24hours.
Quality control strains
Growth
Colony color
Enterococcus faecalis ATCC29212
PR≥0.7
Brown-black halo
Escherichia coli ATCC25922
inhibited
Absence of brown-black halo
Sorage and Shelf Life
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Precautions
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Waste Disposal
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Other Products
Sircol™ – Insoluble Collagen assay kit
Product Info
Document
Product Info
What is Collagen?
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Types of Collagen
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.
Applications of Collagen
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
How does the Sircol assay detect collagen?
Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.
Overview of the Sircol assay process:
Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.
Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then eluted and measured spectrophotometrically.
Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.
Assay range
100 – 1000 µg/ml
Limit of Detection
100µg/ml
Detection Method
Colorimetric Detection (556nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
Suitable Samples
The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.
*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.
**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.
Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Sircol Insoluble Collagen kit contents:
1. Sircol Dye Reagent (1x110ml)
2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)
3. Acid-Salt Wash Reagent (1x20ml)
4. Fragmentation Reagent (1x110ml)
5. Alkali Reagent (1x110ml)
6. 2ml screw-cap tubes for preparation of samples.
7. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.
Rapid and efficient removal of abundant proteins from serum and plasma samples
Process 10 samples in 30 minutes
Allows for visualization of low abundance proteins
Convenient and affordable spin column protocol
Generic protocol allows for depletion from human and other animal samples
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
This kit provides a fast and simple procedure for the effective depletion of major serum proteins including albumin, α-antitrypsin, transferrin and haptoglobin from serum and plasma samples. Such abundant proteins usually obscure less abundant proteins in gel electrophresis making them difficult to visualize and recover. The kit reduces sample complexity to allow for convenient investigation of less abundant proteins. In other applications such as mass spectroscopy, fractionating abundant proteins improves the resolution of less abundant proteins. The kit is ‘generic’ in that it is not antibody-based and can thus be used to deplete abundant proteins from any starting material – human or various other animals – making it more versatile, flexible and affordable than other kits. Resulting proteins are ready for a range of applications including 2D gel electrophoresis, SDS-PAGE, Mass Spectrometry, Protein microarrays, DIGE (Difference Gel Electrophoresis) and more.
Storage Conditions All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
