Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Detail
Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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Sircol™ – Insoluble Collagen assay kit
Product Info
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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.
Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 2 years under recommended storage conditions
Analyte:
Glycerol
Assay Format:
Spectrophotometer, Microplate, Auto-analyser
Detection Method:
Absorbance
Wavelength (nm):
340
Signal Response:
Increase
Linear Range:
1.0 to 35 µg of glycerol per assay
Limit of Detection:
0.37 mg/L
Reaction Time (min):
~ 7 min
Application examples:
Wine (and grape juice), beer, spirits, vinegar, marzipan, fruit juices, soft drinks, toothpaste, honey, tobacco, paper (and cardboard), cosmetics, pharmaceuticals, soap and other materials (e.g. biological cultures, samples, etc.).
Method recognition:
Novel method
The Glycerol GK test kit is a simple, reliable and accurate method for the measurement and analysis of glycerol in beverages, foodstuffs and other material. Based on use of ADP-glucokinase and increase in absorbance on conversion of NAD+ to NADH.
Note for Content: The number of manual tests per kit can be doubled if all volumes are halved. This can be readily accommodated using the MegaQuantTM Wave Spectrophotometer (D-MQWAVE).
Extended cofactors stability. Dissolved cofactors stable for > 1 year at 4oC.
Novel tablet format for increased stability
Very competitive price (cost per test)
All reagents stable for > 2 years as supplied
Very rapid reaction
Positive reaction (assay proceeds with an increase in absorbance)
Mega-Calc™ software tool is available from our website for hassle-free raw data processing
Standard included
Suitable for manual, microplate and auto-analyser formats
Document
The Glycerol GK test kit is a simple, reliable and accurate method for the measurement and analysis of glycerol in beverages, foodstuffs and other material. Based on use of ADP-glucokinase and increase in absorbance on conversion of NAD+ to NADH.
Campylobacter jejuni is a curved, rod-shaped and microaerophillic gram negative bacterium. It is one of the most common causal agents of gastroenteritis with diarrhea as the main symptom. While infection of C. jejuni is seldom life-threatening, it is considered one of the most common food-borne bacteria with over 2 million people infected per year in US alone. Infection of C. jejuni usually results from consumption of poorly prepared food including undercooked meat (particularly poultry), untreated water or raw unpasteurized milk. Traditional identification of C. jejuni involves culturing, however the microaerophilic characteristic of this bacterium make the enrichment process laborious and costly.
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.