| Description | The Cell Cycle and Apoptosis Kit (Cell Cycle and Apoptosis Kit) is a test kit that uses the classic propidium iodide (PI) staining method to analyze the cell cycle and apoptosis. Propidium iodide is a fluorescent dye for double-stranded DNA. When propidium iodide binds to double-stranded DNA, The Cell Cycle and Apoptosis Kit (Cell Cycle and Apoptosis Kit) is a test kit that uses the classic propidium iodide (PI) staining method to analyze the cell cycle and apoptosis. Propidium iodide is a fluorescent dye for double-stranded DNA. When propidium iodide binds to double-stranded DNA, it can produce fluorescence, and the fluorescence intensity is proportional to the content of double-stranded DNA. After the DNA in the cells is stained with propidium iodide, the DNA content of the cells can be measured using a flow cytometer, and then based on the distribution of DNA content, the cell cycle and apoptosis can be analyzed. After staining with propidium iodide, if the fluorescence intensity of G0/G1 phase cells is 1, then the theoretical fluorescence intensity of G2/M phase cells containing double genomic DNA is 2, and the fluorescence intensity of S phase cells undergoing DNA replication is between 1 and 2. Apoptotic cells, due to the condensation of the cell nucleus and the fragmentation of DNA (DNA fragmentation), cause some genomic DNA fragments to be lost during staining, so the iodine staining of apoptotic cells presents a significantly weak color, that is, the fluorescence intensity is less than 1, and a so-called sub-G1 peak appears on the fluorescence graph of flow cytometry, which is the peak of apoptotic cells. When cells undergo apoptosis, due to the concentration of cytoplasm and chromatin, apoptotic bodies are produced, causing changes in the light scattering properties of the cells. In the early stage of cell apoptosis, the ability of the cell to forward-angle light scattering significantly decreases, while the ability of lateral light scattering increases or remains unchanged. In the late stage of cell apoptosis, the signals of forward and lateral light scattering both decrease, so the changes in cell light scattering can be measured using a flow cytometer to observe the apoptosis situation. This kit is usually applied to the detection of the cell cycle and apoptosis of cultured adherent or suspended cells. If used for the cell cycle and apoptosis detection of cells from tissues, the tissue must be digested into a single-cell state before detection can be performed. P1373478Component20 T50 T100 TStorageQuantity Per TestP1373478ADyeing buffer solution10 mL25 mL50 mL-20℃.500 µL per 1 × 10⁶ cells.P1373478BPropidium Iodide staining solution (20X)100 µL250 µL500 µL-20℃.Store in the dark.5 µL per 1 × 10⁶ cells.P1373478CRNase A (50X)0.2 mL0.5 mL1 mL-20℃.10 µL per 1 × 10⁶ cells.Note: The recommended number of cells to stain per test is 1 × 10⁶ cellsInstructions for use1.Collect cells and wash twice with ice-cold PBS; resuspend in ice-cold PBS at a density of 0.1–1.0 × 10⁷ cells/mL.2.In the cell suspension, anhydrous ethanol was added dropwise while shaking until the final ethanol concentration reached 70%.3.Mix gently and fix overnight at 4 °C.4.Wash cells twice with ice-cold PBS, then resuspend in dyeing buffer solution of 2 × 10⁶ cells/mL.5.Add 5 µL of Propidium iodide and 10 µL RNase A to 500 µL of the cell suspension.6.Resuspend cells gently and thoroughly, then incubate at room temperature for 30 min in the dark.7.Analyze samples by flow cytometry.Note: Please bring your own PBS and absolute ethanol.Precautions 1. Fluorescent dyes all have the problem of quenching. Please try to avoid light during storage and use to slow down fluorescence quenching. 2. Iodophor is irritating to the human body. Please take appropriate protective measures. 3. Iodophor is a known mutagen, so this solution needs to be treated with activated carbon before being discarded. 4. When fixing cells with 70% ethanol, make sure it is thorough; otherwise, uneven staining will result in unclear or inaccurate results. It is recommended to fix the cells overnight at -4℃ with 70% ethanol. 5. After cell fixation, ensure that the cells are in a single-cell suspension; cell adhesion may affect our results. 6. For your safety and health, please wear laboratory coats and disposable gloves during operation. 7. This product is only for scientific research and cannot be used for clinical diagnosis or treatment, nor for food or medicine. It must not be stored in ordinary residences. 8. For your safety and health, please wear laboratory coats and disposable gloves during operation... Read More | Products B669892Component50 TStorageB669892ABuffer RCL3×260 mL2-8℃B669892BBuffer GR25 mLRTB669892CBuffer GL25 mLRTB669892DBuffer GW1 (concentrate)13 mLRTB669892EBuffer GW2 (concentrate)15 mLRTB669892FBuffer GE15 mLRTB669892GProteinase K50 mgRTB669892HProteinase K Storage Buffer5 Products B669892Component50 TStorageB669892ABuffer RCL3×260 mL2-8℃B669892BBuffer GR25 mLRTB669892CBuffer GL25 mLRTB669892DBuffer GW1 (concentrate)13 mLRTB669892EBuffer GW2 (concentrate)15 mLRTB669892FBuffer GE15 mLRTB669892GProteinase K50 mgRTB669892HProteinase K Storage Buffer5 mLRTB669892ISpin Columns DL with Collection Tubes50 setsRTProductsThis kit is suitable for the extraction of total DNA, including genomic DNA, mitochondrial DNA and viral DNA, from fresh or frozen whole blood (blood samplestreated with anticoagulants such as citrate, EDTA or heparin), plasma, serum, haematocrit brown and yellow layers, bone marrow, cell-free body fluids, etc. Theproduct can process 1-5 ml of whole blood, and can be purified to obtain sizes rangingfrom 100bp to 50kb. The purified DNA is of high yield and good quality, with maximumremoval of proteins, pigments, lipids and other inhibitory impurities, and can bedirectly used in PCR, fluorescence quantitative PCR, enzyme digestion and SouthernBlot.Self-contained reagent: anhydrous ethanol.Pre-experiment Preparation and Important Notes1. Add 5ml Proteinase K Storage Buffer to Proteinase K to dissolve it, and storeit at -20℃. Do not leave the prepared Proteinase K at room temperature for a longtime, and avoid repeated freezing and thawing to avoid affecting its activity.2. Repeated freezing and thawing of the sample should be avoided, as this may resultin smaller DNA fragments and a decrease in the amount of extracted DNA. 3.This kit can extract up to 1-5 ml of whole blood samples, if you need to extracta large number of blood samples, please use the blood genome non-column extractionkit. 4. Anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to theinstructions on the label of the reagent bottle before first use.5. Please check Buffer GL for crystallization or precipitation before use, if thereis any crystallization or precipitation, please put it in 56℃water bath to re-dissolve.6. If the downstream experiments are sensitive to RNA contamination, 4µl of DNaseFree RNase A (100mg/ml) can be added, RNase A is not provided in the kit, and canbe ordered separately from our company if needed.7. The Buffer RCL in the kit cannot be used further after turbidity.procedure1. Add 1-5 ml of blood sample to a centrifuge tube (supplied) and add 3 times thevolume of Buffer RCL and gently vortex or invert to mix.2. Centrifuge at 3000 rpm (~900 x g) for 10 minutes and carefully aspirate thesupernatant.3. Add 400 µl Buffer GR to the precipitate and resuspend the precipitate. Note: If the downstream assay is sensitive to RNA, add 4 µl of RNase A (100 mg/ml)solution, shake for 15 seconds, and leave at room temperature for 5 minutes.4. For 1-2 ml blood sample extraction, add 40µl Proteinase K to the above solutionand mix well; for 2-5 ml blood sample extraction, add 100µl Proteinase K to theabove solution and mix well.5. Add 400 µl of Buffer GL, mix upside down 15 times, and vigorously vortex andshake for at least 1 minute. Note: Do not add Proteinase K directly to Buffer GL.6. Incubate at 70°C for 10 minutes, during which time mixing was inverted severaltimes.Note: 1) If the solution is not completely clear, add appropriate amount of Proteinase K and incubate. Extend the incubation time until the solution is completely clear. 2) The yield of DNA has been maximized by 10 minutes of incubation, and continuedprolongation of the incubation time has no effect on DNA yield or purity.7. Add 400 µl of anhydrous ethanol and mix upside down 10 times. Centrifuge brieflyto concentrate the liquid on the walls and cap to the bottom of the tube.8. Add all of the solution obtained in the previous step to the Spin Columns DL inthe collection tube. If the solution cannot be added all at once, transfer it severaltimes. centrifuge at 12,000 rpm (~13,400 x g) for 1 minute, pour off the waste liquidfrom the collection tube, and put the column back into the collection tube.9. Add 500 µl of Buffer GW1 to the adsorption column (check that anhydrous ethanolis added before use), centrifuge at 12,000 rpm for 1 minute, pour off the waste liquidin the collection tube, and put the adsorption column back into the collection tube.Note: It is recommended that step 9 be repeated if the sample being extracted isthe blood genome of a species such as mice or monkeys from which hemoglobin isdifficult to remove.10. Add 500 µl Buffer GW2 to the adsorption column (check that anhydrous ethanolis added before use), centrifuge at 12,000 rpm for 1 minute, pour off the waste liquidin the collection tube, and put the adsorption column back into the collection tube.Note: Step 10 can be repeated if further DNA purity is required.11. Centrifuge at 12,000 rpm for 2 minutes and pour off the waste liquid in thecollection tube. Leave the adsorption column at room temperature for several minutesto dry thoroughly. Note: The purpose of this step is to remove residual ethanol from the adsorptioncolumn, which can interfere with subsequent enzymatic reactions (digestion, PCR,etc.)12. Place the adsorption column in a new centrifuge tube, add 50-200 µl of BufferGE or sterilized water to the middle of the adsorption column overhanging the column,leave it at room temperature for 2-5 minutes, centrifuge at 12,000 rpm for 1 minute,collect the DNA solution, and store the DNA at -20℃.Note: 1) If the downstream experiment is sensitive to pH or EDTA, you can use sterilized water for elution. The pH of the eluent has a great influence on theelution efficiency, if water is used as the eluent should ensure that its pH is7.0-8.5 (you can use NaOH to adjust the pH of the water to this range), and the elutionefficiency is not high when the pH is lower than 7.0.2) Incubation at room temperature for 5 minutes prior to centrifugation increasesyield.3) Re-elution with an additional 50-200 µl Buffer GE or sterilized water can increase the yield.4) If the final concentration of DNA is to be increased, the DNA eluate obtainedin step 12 can be re-spiked onto the adsorbent membrane and centrifuged at 12,000rpm. 1min; if the elution volume is less than 200µl, the final concentration of DNA canbe increased, but the total yield may be reduced. If the amount of DNA is less than1 µg, elution with 50 µl Buffer GE or sterilized water is recommended.5) Because DNA preserved in water is subject to acidic hydrolysis, for long-termstorage, it is recommended that it be eluted with Buffer GE and stored at -20℃... Read More | Format:2-ComponentEnzyme:Horseradish peroxidase | DescriptionCobalt is a transition metal that serves as a trace dietary mineral for all multicellular organisms. Cobalt is an important cofactor for the Vitamin B12class of compounds where it occupies the center of the vitamin B12corrin ring. Cobalt can also be coordinated in the active site of the DescriptionCobalt is a transition metal that serves as a trace dietary mineral for all multicellular organisms. Cobalt is an important cofactor for the Vitamin B12class of compounds where it occupies the center of the vitamin B12corrin ring. Cobalt can also be coordinated in the active site of the non-corrin containing metalloenzyme methionine aminopeptidase.Suitability: Suitable for quantitating cobalt concentrations in a variety of samplesPrinciple: The Cobalt Assay kit provides a simple and direct procedure for measuring cobalt in a variety of samples. In this assay, cobalt reacts with 2-mercaptoethanol under basic conditions to form a complex with a strong absorbance at 475 nm. Interference from the metal ions Fe2+, Cu2+, Ni2+, Zn2+, and Mn2+is <10% at this wavelength. This assay gives a linear range of 10-50 nmoles of cobalt.}Preparation instructionsSuitable for quantitating cobalt concentrations in a variety of samplesPrincipleThe Cobalt Assay kit provides a simple and direct procedure for measuring cobalt in a variety of samples. In this assay, cobalt reacts with 2-mercaptoethanol under basic conditions to form a complex with a strong absorbance at 475 nm. Interference... Read More | DescriptionThe 1 µm Coupling Kit makes conducting immunoprecipitation and biomolecule separation easier and more flexible. The Kit contains AnteoBind™activated 1 µm magnetic particles that give you increased antibody binding capacity and functionality, while the included blocking DescriptionThe 1 µm Coupling Kit makes conducting immunoprecipitation and biomolecule separation easier and more flexible. The Kit contains AnteoBind™activated 1 µm magnetic particles that give you increased antibody binding capacity and functionality, while the included blocking buffer decreases background noise. Reduce reagent preparation time; remove traditional surface preparation steps such as EDC and replace these steps with the 1 µm pre-activated magnetic particles provided. This Kit reduces aggregation and gives you the freedom and ability to develop multifunctional particles for diverse applications, including dual labelling.Binding Capacity and Dispersity:Binding Capacity:> 20 µg IgG/mgMonodispersity:> 90% (by light microscopy determination)Particle based immunoassays, bioseparations and immunoprecipitation... Read More |