Biotin Fluorescence in Situ Hybridization Kit for RNA (Green)
$500.00 - $2,000.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: BFHKRG-100 (for 100T)
Cat. No.: BFHKRG-500 (for 500T)
Description
Biotin Fluorescence in Situ Hybridization Kit for RNA (Green), also known as the Biotin FISH Kit for RNA (Green), is a high-sensitivity detection kit. It uses biotin-labeled oligonucleotide probes (DNA or RNA) to hybridize with target RNA in cells or tissues based on the principle of complementary base pairing. The resulting biotin-labeled nucleic acid probe and target RNA hybrid complex then binds to Alexa Fluor 488-labeled streptavidin (Streptavidin-488), which can subsequently be observed under a fluorescence microscope to determine the intracellular localization of the target RNA.
Streptavidin is a tetrameric protein that can simultaneously bind with high specificity to four biotin molecules. This natural biomolecular interaction system, known as the biotin-streptavidin system, is characterized by high affinity, sensitivity, specificity, and stability. It has been widely applied in various detection systems, such as in situ hybridization, immunodetection, DNA hybridization assays, immunohistochemistry, flow cytometry, and the purification and characterization of target molecules. This system aids in signal amplification and enhances detection sensitivity.
Streptavidin-488 can specifically bind to the biotin-labeled nucleic acid probe and target RNA hybrid complex. Alexa Fluor 488 is a commonly used green fluorescent probe with a maximum excitation wavelength of 493 nm and a maximum emission wavelength of 519 nm, producing very bright green fluorescence observable under a fluorescence microscope.
Components
- Proteinase K (20mg/ml)
- Acetylation Solution A
- Acetylation Solution B
- Hybridization Solution
- Yeast RNA (100X)
- Streptavidin Buffer (5X)
- Streptavidin-488
- DAPI (5mg/ml)
- Wash Buffer (20X)
Features
- This kit has low background and high sensitivity. It includes specialized acetylation reagents and a separate blocking reagent, Yeast RNA (100X), which effectively reduces background and enhances detection sensitivity.
- Wide range of applicable samples. This kit can be used for target RNA localization analysis in various samples, including tissues fixed with paraformaldehyde or formalin, paraffin-embedded or frozen sections, and cells. It is suitable for detecting sample heterogeneity, high-sensitivity single-cell detection of non-coding RNA (including miRNA and lncRNA), tracking genomic RNA, and high-throughput single-cell quantitation of target RNA.
- Provides single-cell spatial distribution information for target RNA. Gene expression analysis typically employs technologies such as transcriptome sequencing, qPCR, Northern blotting, and gene microarray chips. While these methods offer extensive information, they often mask single-cell gene expression differences, particularly the spatial distribution of RNA within cells. This kit allows for the specific and highly sensitive revelation of the spatial distribution of gene expression at the single-cell level.
- Compatible with subsequent fluorescence staining. The kit exhibits good fluorescence compatibility. In addition to the specific hybridization fluorescence signal, it can be used with fluorescent dyes such as DAPI, Hoechst 33342, Hoechst 33258, and Propidium Iodide.
Storage
Store at -20ºC, valid for one year. Proteinase K (20mg/ml) and Yeast RNA (100X) should be stored at -20ºC. Hybridization Solution, Streptavidin-488, and DAPI should be stored at -20ºC, protected from light. Acetylation Solution A, Acetylation Solution B, Streptavidin Buffer (5X), and Wash Buffer (20X) can be stored at 4ºC or room temperature.
Note
- It is recommended to use fresh samples for FISH detection whenever possible. For preserved samples, RNA typically degrades over time, which can affect the FISH detection results.
- Reagents needed:
- Ultrapure Water (DNase/RNase-Free, Sterile) or DEPC Water
- 4% Paraformaldehyde Fixative Solution
- PBS, pH 7.3 (DNase, RNase & Protease Free, Sterile)
- BSA (Fatty Acid & IgG Free)
- Hydrochloric Acid
- Formamide Deionized
- Anti-Fade Mounting Medium
- Poly-L-Lysine (as needed)
- Coverslips (as needed)
- Reagents for Frozen or Paraffin Sections (as needed)
- Biotin-labeled Probe for Target RNA (must be purchased or synthesized separately)
- For routine detection of mammalian cells or tissue samples, it is usually effective to add Yeast RNA to the Hybridization Solution before use to reduce background. In cases where Yeast RNA might interfere, it can be omitted.
- It is advisable to aliquot and store Hybridization Solution and Streptavidin-488 protected from light.
- Once the pre-hybridization temperature is determined, the temperature for subsequent probe pretreatment, hybridization, and washing should be consistent.
- All reagents must be fully thawed and mixed before use.
- Due to the involvement of RNA procedures, it is essential to strictly follow RNA handling protocols to avoid RNase contamination. Pipette tips, centrifuge tubes, and related equipment and solutions used in the experiment must be RNase-free. Glassware, plasticware, instruments, and work surfaces should also be treated to be RNase-free whenever possible.
- This product is for scientific research use by professionals only and is not intended for clinical diagnosis or treatment, nor for use in food or drugs, and should not be stored in a household environment.
- For your safety and health, wear a lab coat and disposable gloves when handling.
Related: Fluorescence in Situ Hybridization Kit for RNA
Only for research and not intended for treatment of humans or animals
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