| Description | Product Introduction:1. The Biotin Assay Blocking Kit is a kit primarily designed for blocking tissue and cell samples to significantly reduce background during detection processes such as immunohistochemistry (IHC), immunocytochemistry (ICC), immunofluorescence (IF), and fluorescence in situ Product Introduction:1. The Biotin Assay Blocking Kit is a kit primarily designed for blocking tissue and cell samples to significantly reduce background during detection processes such as immunohistochemistry (IHC), immunocytochemistry (ICC), immunofluorescence (IF), and fluorescence in situ hybridization (FISH). These detection methods are based on biotin and streptavidin or avidin, including the SABC (Streptavidin-Biotin Complex) method and ABC (Avidin-Biotin Complex) method.2. This kit contains two types of blocking buffers: Biotin Blocking Buffer (100X): Contains streptavidin and other components, which can fully bind to and block endogenous biotin, and reduce non-specific binding to labeled streptavidin in subsequent steps. Streptavidin/Biotin-Binding Blocking Buffer (100X): Contains biotin, which can fully block endogenous biotin-binding proteins and the streptavidin bound during the previous blocking step. Through this two-step blocking process, the goal of significantly reducing background can be achieved.3. This kit is suitable for blocking frozen sections and paraffin sections, and also applicable for blocking cultured cells.Product Features:1. Easy to Use – Can be performed simultaneously with the blocking step of conventional immunostaining, significantly saving blocking time.2. Stable Performance – Effectively reduces non-specific staining and improves detection sensitivity.Product Components and Storage Conditions:Product numberComponent200TStorageB1209118ABiotin Blocking Buffer (100×)200 µl-20ºCB1209118BStreptavidin/Biotin-Binding Blocking Buffer (100×)200 µl-20ºCOperating Steps:1. For immunostaining assays such as IHC, ICC, and IF:a. Dilution of blocking buffers: Based on the required volume, dilute Biotin Blocking Buffer and Streptavidin/Biotin-Binding Blocking Buffer to 1X using an appropriate conventional blocking buffer.b. After thorough washing of the sections and before primary antibody incubation, add an appropriate amount of Biotin Blocking Buffer and incubate for 10-30 minutes. Incubation for 10 minutes is recommended when diluted with a rapid immunostaining blocking buffer; incubation for 30 minutes is recommended when diluted with a regular blocking buffer.c. Wash the samples with washing buffer 3 times, 5 minutes each.d. Add an appropriate amount of Streptavidin/Biotin-Binding Blocking Buffer and incubate for 10-30 minutes. Incubation for 10 minutes is recommended when using a rapid immunostaining blocking buffer; incubation for 30 minutes is recommended when using a regular blocking buffer.e. Wash the samples with washing buffer 3 times, 5 minutes each.f. Proceed with subsequent steps such as primary antibody incubation according to conventional procedures.2. For in situ hybridization (FISH) detection:a. Dilution of blocking buffers: Based on the required volume, dilute Biotin Blocking Buffer and Streptavidin/Biotin-Binding Blocking Buffer to 1X using an appropriate diluent. The washing buffer for in situ hybridization can be used for dilution.b. Before hybridization with biotin-labeled probes, add an appropriate amount of Biotin Blocking Buffer and incubate for 15-30 minutes.c. Wash the samples with washing buffer 3 times, 5 minutes each.d. Add an appropriate amount of Streptavidin/Biotin-Binding Blocking Buffer and incubate for 15-30 minutes.e. Wash the samples with washing buffer 3 times, 5 minutes each.f. Proceed with subsequent steps such as hybridization with biotin-labeled probes according to conventional in situ hybridization detection procedures.Precautions:1. This kit can also perform the above-mentioned blocking steps after the primary antibody incubation is completed and the sample is washed 3-4 times with washing buffer, followed by subsequent steps such as secondary antibody incubation.2. This kit can simultaneously perform biotin-related blocking and conventional blocking, eliminating the need for additional conventional immunostaining blocking steps.3. For your safety and health, please wear a lab coat and disposable gloves during operation... 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 | Product content: G665990Component200 TStorageG665990ABuffer PG100 mLRTG665990BBuffer PS60 mLRTG665990CBuffer PW (concentrate)50 mLRTG665990DBuffer EB30 mLRTG665990ESpin Columns DM with Collection Tubes200 EART Product Introduction:This kit uses a new silicon-based plasma membrane technology and Product content: G665990Component200 TStorageG665990ABuffer PG100 mLRTG665990BBuffer PS60 mLRTG665990CBuffer PW (concentrate)50 mLRTG665990DBuffer EB30 mLRTG665990ESpin Columns DM with Collection Tubes200 EART Product Introduction:This kit uses a new silicon-based plasma membrane technology and reagent formulation. Through the unique centrifugal adsorption column and the DNA washing elution step, 100 bp-10 kb DNA fragments can be recovered and purified from ordinary or low melting point agarose gel. The sol speed is fast and the recovery rate is high. The sol solution contains a pH indicator, which can be used to determine whether the sol recovery has reached the optimal state based on its color. Each adsorption column can adsorb up to 10 µ G DNA, while effectively removing impurities such as primers, enzymes, mineral oil, and agarose. The purified and recovered DNA has high purity and concentration, good integrity, and can be directly used for molecular biology experiments such as sequencing, linking and transformation, labeling, and in vitro transcription.Self prepared reagents: anhydrous ethanol, isopropanol.Preparation and important precautions before the experiment:1.Before the first use, anhydrous ethanol should be added to the Buffer PW according to the instructions on the reagent bottle label.2. Before use, please check the Buffer PG. If crystallization or precipitation occurs, it can be left in a 37 ℃ water bath for 3-5 minutes to restore clarity.3. It is best to use a new electrophoresis buffer during electrophoresis to avoid affecting the electrophoresis and recovery efficiency; The following experiment requires high requirements, please use TAE electrophoresis buffer as much as possible.4.When cutting glue, the UV irradiation time should be as short as possible to avoid damage to DNA.5. The recovery rate is related to the initial amount of DNA and the elution volume. The smaller the initial amount, the smaller the elution volume, and the lower the recovery rate.6. Preheat the water bath to 50 ℃.7. Buffer PG contains a pH indicator. When the pH is ≤ 7.5, the color of the solution is yellow, and DNA can effectively bind to the membrane. When the pH is too high, the color of the solution turns orange red and purple, which needs to be adjusted.8. All centrifugation steps can be performed at room temperature.Operation steps:1. Cut the single purpose DNA strip from the agarose gel (try to cut the excess), put it into a clean centrifuge tube (self prepared), and weigh and calculate the weight of the gel (record the weight of the centrifuge tube in advance).Attention: If the volume of the adhesive block is too large, it can be cut into small pieces.2. Add one time of the volume of Buffer PG (if the gel weighs 100 mg, its volume can be regarded as 100 µ l. And so on.3.50 ℃ water bath and gently invert the centrifuge tube every 2-3 minutes until the sol turns yellow to ensure full dissolution of the gel block. If there are still unsolved glue blocks, you can add some more sol solution or continue to let it stand for a few minutes until the glue blocks are completely dissolved.Note: 1) After the gel is completely dissolved, the gel solution is yellow, and subsequent operations can be carried out; If the glue solution is orange red or purple, 10-30 can be added to the glue solution µ 3 M sodium acetate (pH 5.0), adjust the color of the solution to yellow before proceeding with subsequent operations.2) After the gel block is completely dissolved, it is best to lower the temperature of the gel solution to room temperature before loading the column. The adsorption column has a weaker ability to bind DNA at higher temperatures.4. (Optional step) When the recovered fragment is less than 300 bp, add 1/2 of the gel volume of isopropanol, and mix it upside down (if the gel weighs 100 mg, add 50 µ Isopropanol of L.5. Column balance: Add 200 to the spin columns DM that have been loaded into the collection tube µ Centrifuge at 13000 rpm (~16200 × g) for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.6. Add the solution obtained from steps 3 or 4 to the adsorption column that has been loaded into the collection tube, let it stand at room temperature for 2 minutes, centrifuge at 13000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column in the collection tube.Attention: The volume of the adsorption column is 750 µ l. If the sample volume is greater than 750 µ L can be added in batches.7. Add 450 to the adsorption column µ LBuffer PW (please check if anhydrous ethanol has been added before use), centrifuge at 13000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column in the collection tube.Note: If purified DNA is used for salt sensitive experiments (such as flat end ligation or direct sequencing), it is recommended to add Buffer PW and let it stand for 2-5 minutes before centrifugation.8. Repeat step 7.9.13000 rpm for 1 minute and discard the waste liquid from the collection tube.Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.).10. Place the adsorption column into a new 1.5 ml centrifuge tube (provided by oneself), and add 50 drops to the middle position of the adsorption membrane in the air µ L Buffer EB, leave at room temperature for 2 minutes. Centrifuge at 13000 rpm for 1 minute and collect DNA solution- Store DNA at 20 ℃.Attention:1) To improve the recovery of DNA, the solution obtained by centrifugation can be re dropped onto the adsorption column, left at room temperature for 2 minutes, and centrifuged at 13000 rpm for 1 minute.2) The elution volume should not be less than 30 µ l. A small volume will affect the recovery efficiency.3) When recovering DNA fragments larger than 10 kb, Buffer EB should be preheated in a 50 ℃ water bath to increase recovery efficiency.Note: This reagent kit is also suitable for the purification and recovery of PCR products. Add an equal volume of Buffer PG to the PCR reaction solution and mix thoroughly (for small fragments with a recovery of less than 150bp, the solution volume can be increased to three times to improve the recovery rate). Follow step 5 above for further operations... Read More | Product introduction:Reporter gene detection is an important tool for analyzing the interaction between potential cis elements (such as promoters, enhancers and silencers) and trans acting factors in the flanking region of structural genes in the field of modern molecular biology. Firefly Product introduction:Reporter gene detection is an important tool for analyzing the interaction between potential cis elements (such as promoters, enhancers and silencers) and trans acting factors in the flanking region of structural genes in the field of modern molecular biology. Firefly luciferase is widely used in gene regulation and drug screening. Firefly luciferase is a protein with a molecular weight of about 61 KD. In the presence of ATP, magnesium ions and oxygen, it can catalyze the production of oxyluciferin from luciferin. In the process of luciferin oxidation, it will produce a light signal. The optical signal of this kit is a kind of instantaneous light, which needs to be detected immediately after adding the working solution. The half-life of optical signal is about 5 min.Instruction:1.Working fluid configuration ( 1 ) Restore all components to room temperature. ( 2 ) The component B ( stock solution ) was fully diluted with component A to prepare a 0.2 mg / mL firefly luciferase working solution, which was vortexed and shaken to ensure full mixing. Note : The firefly luciferase working solution cannot be repeatedly frozen and thawed. If the dosage of a single experiment is small, it is recommended to subpackage according to a single dosage. At room temperature, the activity decreased by about 10 % after the working solution was configured for 3 h, and the activity decreased by about 25 % after 5 h. 2.chemiluminescence value detection ( 1 ) The cell culture plate was taken out from the incubator and incubated at room temperature for 20 min to restore it to room temperature ( 22-25 ° C ). ( 2 ) Add the same volume of firefly luciferase working solution with the medium to the culture plate and mix well. ( 3 ) Incubation at room temperature for 5 min. Note : The incubation time can be adjusted according to cell type and cell number. ( 4 ) The values were read by multifunctional microplate reader or chemiluminescence instrument ( instrument parameters : the determination time was 10 s, the determination interval was 2 s ).Matters needing attention:1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments. 2. the strongest wavelength of bioluminescence catalyzed by firefly luciferase is 560 nm. 3. to prevent interference between holes, it is recommended to use white opaque orifice plate.Recommendation:Component B is recommended to use sterile water in advance to configure 2 mg / mL storage solution, A component and B component configured as storage solution, and small batch packaging according to the experimental requirements. The detection working fluid is recommended to be used now to avoid repeated freezing and thawing. Component:One-Step Firefly Luciferase Assay Buffer;D-Luciferin Scope of application:Mainly used for ADCC detection... Read More | DescriptionMaterials included in the kit are designed to be used with the Hy-Energy′s PCTPro-2000 System. They also can be used for demonstration purposes and as standards during the development of novel hydrogen storage and battery materials |