Principle: Peptone provide carbon and nitrogen sources to meet the needs of bacterial growth; lactose are fermentable sugars; selenite, sodium hydrogen inhibit Gram-positive bacteria and gram-negative enterobacteria most non-Salmonella; phosphate-buffered agent; L- cystine as a reducing agent.
How to use: 1. Weigh 23g of the product , adding 1 L of distilled or deionized water , heated to boiling stirring until completely dissolved, dispensing flask, cooled to room temperature . 2. Pipette 10mL of pre-enrichment sample broth or 25mL and transferred species in the liquid sample flask in a sterile environment. 3.Place into incubator, cultured at 36 ± 1 for 18-24h. 4. Observe the results.
Quality control: Quality control strains were inoculated ,and cultured at 36 ± 1 for 18-24h ,results show as follows: strain name strain code growth feature Salmonella typhi CMCC (B) 50071 good red, cloudy Salmonella typhimurium CMCC (B) 50115 good red, cloudy Escherichia coli ATCC25922 — remain unchanged
Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
Mycobacterium tuberculosis is a pathogenic bacterial species belonging to the genus Mycobacterium, and is the causative agent of tuberculosis. Tuberculosis (TB) is a multifaceted disease and challenging public health problem in both industrialized and developing countries. According to the WHO, 8.8 million active cases of TB are diagnosed each year and of these, almost 2 million die. Once thought to be under control or even close to extinction, TB infection levels are rising and the threat is compounded by new, virulent and drug-resistant strains. Although most cases occur in the developing world (22 countries accounting for 80% of all global cases), increasing population mobility combined with facility of transmission means that no country is immune from the resurgence of TB. TB control programs are currently facing a number of constraints. Worldwide, fewer than 25% of all tuberculosis cases are detected. Of utmost concern is the absence of a timely and accurate test for the diagnosis of mycobacterial disease. Early diagnosis is crucial for the prevention of further spread of the disease.
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
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 SOLUBLE forms of collagen. Chose the Sircol Insoluble collagen kit if you need to analyse INSOLUBLE 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 Sircol 2.0 detect collagen?
The Sircol 2.0 dye reagent includes Sirius Red, a linear anionic dye with sulfonic acid side chains. This reagent is specially formulated to bind to the Gly-X-Yn helical structure of soluble collagen under assay conditions.
*The improved formulation of Sircol 2.0 dye enables a greater degree of dye-collagen specificity (compared to our previous S1000 assay kit).
Overview of the Sircol 2.0 assay process:
Step 1. Prepared samples are placed in the wells of the assay microplate, together with Sircol Dye Reagent. After 30 minutes mixing, any collagen-dye complexes will form as a precipitate. These are collected on the base of the microplate wells by centrifugation.
Step 2. Unbound dye is removed by gentle aspiration, followed by a rinse with Plate Wash Reagent.
Step 3. Following further centrifugation, collagen-bound dye is eluted by incubation with a Dye Release Reagent. Eluted dye is detected ‘in-situ’ by spectrophotometric analysis of the microplate at 556nm.
Step 4. The collagen content of unknown samples can be quantified by comparison against a calibration curve, prepared using the Collagen Reference Standard supplied with the kit.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
Colorimetric Detection (556nm) (Endpoint), Requires a microplate centrifuge.
Measurements per kit
96 in total (allows a maximum of 41 samples to be run in duplicate alongside a standard curve).
Suitable Samples
Soluble* collagens from mammalian**:
In-vivo: Tissues, cartilages and fluids.
In-vitro: Extracellular matrices / Conditioned media from 2D/3D culture environments.
The straightforward sample processing and analysis of Sirco 2.0 make it a good alternative to conventional hydroxyproline analysis.
*Prior salt/acid/acid-pepsin extraction may be necessary to release soluble collagen.
**Sircol 2.0 is primarily designed for use with in-vivo / in-vitro samples of mammalian origin. Collagens originating from other taxonomic groups and kingdoms can also be analysed. See note on p6 of manual for further information.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a microplate centrifuge* (see note below), 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.
*As a minimum, we recommend that the centrifuge can centrifuge a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
Sircol 2.0 kit contents:
1. Dye Reagent (1x20ml)
2. Collagen Reference Standard (1x5ml, 200µg/ml of soluble Bovine collagen)
3. Plate Wash Reagent (1x28ml)
4. Collagen Concentration Reagent (1x25ml)
5. Neutralisation Reagent (1x8ml)
6. Dye Release Reagent (1x25ml)
7. Assay Microplate (1×96-wells)
8. Microplate Seals (6x)
9. Documentation (QuickStart Guide / Manual / Certificate of Analysis)
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details. This kit requires the use of a microplate centrifuge, capable of centrifuging a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
Document
Experience user-friendly detection & measurement of Soluble Collagen with Sircol™ 2.0! Our latest kit simplifies collagen quantification within in-vivo / in-vitro samples. Sircol 2.0 offers enhanced sensitivity and accuracy compared to our previous Sircol kit.
Thirteen discrete fragments ranging from 300 bp to 10000 bp
Higher intensity reference band at 5000 bp
The HighRanger 1 kb DNA Ladder is prepared to ensure quality and batch-to-batch consistency. This Ladder contains thirteen discrete fragments ranging from 300 bp to 10,000 bp with a higher intensity reference band at 5000 bp. This Ladder is ideal for quick sizing of PCR products and restriction digests.
Contents 1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).
HighRanger 1kb DNA Ladder (Cat# 11900) – 100 loads
Ladder Properties: • Thirteen discrete bands, ranging from 300 bp to 10000 bp • Higher intensity band at 5000 bp for easy reference
Fragment
Size (bp)
Mass (ng)
1
10000
58
2
8000
53
3
6000
46
4
5000
67
5
4000
56
6
3000
47
7
2500
41
8
2000
34
9
1500
29
10
1000
18
11
700
13
12
500
23
13
300
14
Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.
Storage:
This kit is stable for 2 years after the date of shipment.
This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.