Mycoplasma is a common and serious contaminant of cell cultures. Mycoplasma is often not visible and does not respond to antibiotics, and therefore it is a major issue that requires monitoring and early detection. Up to 30% of cell cultures may be contaminated with Mycoplasmas, the main contaminants being the species M. orale, M. laidlawii, M. arginini and M. hyorhinis. These organisms produce a variety of effects on the infected cells in culture, including changes in metabolism growth, viability and morphology, thereby altering the phenotypic characteristics of the host cells. Therefore there is an absolute requirement for routine, periodic assays to diagnose possible Mycoplasma contamination of all cell cultures, particularly continuous or established cell lines.
Mycoplasma TaqMan PCR Kit, 100 reactions
Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
Specific Primer and Probe mix for the pathogen/virus/viroid of interest
Primer and Probe mix
Positive and negative control to confirm the integrity of the kit reagents
Mycoplasma TaqMan PCR Probe/Primer Set and Controls, 100 reactions
Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
Nuclease-free water
Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix
For research use only and NOT intended for in vitro diagnostics.
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. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
Component
Cat. TM33150 (100 preps)
Cat. TM33110 (100 preps)
MDx TaqMan 2X PCR Master Mix
2 x 700 μL
–
Mycoplasma Primer & Probe Mix
280 μL
280 μL
Mycoplasma Positive Control
150 μL
150 μL
Nuclease-Free Water (Negative Control)
1.25 mL
1.25 mL
Product Insert
1
1
Other Products
IST-125 QuickSeal MicroTM Self Adhesive Sealing Film
Product Info
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Product Info
Overview
Low strength adhesive film that is transparent, peelable and suitable for short term storage.
This transparent polyester-based film has a low strength adhesive
It is designed as a low-cost sealing option, and useful for temporary storage and as a cover for applications such as centrifugation
End tabs allow for easy application and removal
This seal is removable without leaving residue on the plate
For all adhesive seals, the best sealing results are achieved using our Hand Roller or KAPS 500 Auto Sealer
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Low strength adhesive film that is transparent, peelable and suitable for short term storage.
Propargyl-PEG14-alcohol is a propargyl linker which enables Click Chemistry with azide compounds to yield stable triazole linkage. The PEG spacer enhances the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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Propargyl-PEG14-alcohol is a propargyl linker which enables Click Chemistry with azide compounds to yield stable triazole linkage. The PEG spacer enhances the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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.
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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.
