Description
Specifications
| Clone | IHC583 |
| Source | Mouse Monoclonal |
| Positive Control | Breast Carcinoma, Urothelial Carcinoma |
| Dilution Range | 1:200 |
GATA3 is a transcription factor important in cell proliferation, development, and differentiation. GATA3 is mostly observed in breast and urothelial carcinomas, and rarely present in other cancers such as endometrial endometrioid adenocarcinoma. Among the breast carcinomas, GATA3 has a lower expression in luminal B subtype breast carcinoma. Studies have found GATA3 expression to be associated with ER (estrogen receptor), PR (progesterone receptor), and Her2 in breast cancer cases. Urothelial carcinomas stain positively for GATA3 in invasive or high grade tumors, therefore Anti-GATA3 is useful for carcinoma diagnosis when breast and bladder are plausible.
| Clone | IHC583 |
| Source | Mouse Monoclonal |
| Positive Control | Breast Carcinoma, Urothelial Carcinoma |
| Dilution Range | 1:200 |
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from FFPE tissue and section samples (with DNase) |
| Applications | RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNase |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | FFPE tissue sample |
| Sample amount | 6mg |
| Yield | 20μg |
| Elution volume | ≥10μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
Kit Contents
| Contents | R414402 | D414403 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Micro Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Buffer DPS | 60 ml | 250 ml |
| Buffer FRL | 15 ml | 60 ml |
| Buffer RLC | 15 ml | 60 ml |
| Buffer RWC* | 10 ml | 50 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| DNase I | 600 µl | 5 x 600 µl |
| DNase Booster Buffer | 1.5 ml | 6 ml |
| Protease Dissolve Buffer | 1.8 ml | 10 ml |
| Proteinase K | 24 mg | 120 mg |
| RNase Free Water | 10 ml | 20 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Experiment Data
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
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).
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.
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.
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).
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.
2 – 200µg/ml or 0.25 – 20µg/ml (using optional Collagen Concentration Protocol)
2 µg/ml
Colorimetric Detection (556nm) (Endpoint), Requires a microplate centrifuge.
96 in total (allows a maximum of 41 samples to be run in duplicate alongside a standard curve).
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.
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.
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.
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.