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We narrowed to 545 results for: SEC

Showing: 261 - 280 of 545 results

  3. Antibodies 101: Validation

    Type
    Blog Post
    ...complex samples such as whole cell lysate or tissue sections. It is important to confirm that your antibody...doi.org/10.1080/19420862.2015.1100787  Taussig MJ, Fonseca C, Trimmer JS (2018) Antibody validation: A view...

  4. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ... you may not find in the materials and methods sections of optogenetics papers. Optogenetics step one...channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function. Nature methods....

  5. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...and corresponding Multi-rAb CoraLite® Plus 647 secondary antibodies. Antibody staining was quantified using...histograms show cells that have only been stained with secondary antibodies, showing the level of background staining...

  6. Deep Dive: qPCR

    Type
    Blog Post
    ...design resources in the References and Resource section. Validation Once designed, your primer and probe...amplicons; if your amplicon is AT-rich or has a secondary structure, you may see a double peak even with...

  7. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...N-glycosylase (UNG) during base excision repair. Adding a second copy of the UNG inhibitor, UGI, increases base ...BE4-Gam; AID, CDA1 and APOBEC3G BE3 variants   A second copy of UGI improves product purity. Gam decreases...

  8. Western Blot

    Type
    Protocol
    ... anti-mouse secondary antibody for primary antibodies raised in a mouse. Procedure Section 1: Lyse cells...incubate with primary antibody Day 2: Incubate with secondary antibody Video Watch this instructional video ...Chemiluminescence substrate Plastic wrap Primary antibody Secondary antibody Deionized water Before Starting Refer... your antibody to determine the optimal dose. Secondary antibodies must match the host species of the ...immediately or store at -80 °C until ready to use. Section 2: Determine the total protein concentration and...to 1X . Boil the samples for 10 min at 100 °C . Section 3: SDS-PAGE Prepare the precast gel as follows:...separation. Gently open the gel with the spatula. Section 4: Dry transfer Soak the gel for 15 min in 20% ...

  9. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ... room temperature or overnight at 4 °C . Section 4: Secondary antibody incubation Carefully remove the...Blocking Day 3: Primary antibody incubation Day 4: Secondary antibody incubation and plate read Tips and Troubleshooting...control antibody HRP-conjugated isotype-specific secondary antibody ELISA 96-well microplate, Corning 9018...Starting Warm reagents to room temperature. Procedure Section 1: Prepare the Antigen Standard and coat the plate...incubate at 37 °C for 30 min , or overnight at 4 °C . Section 2: Block the plate Prepare the wash buffer (0.05%...2 h at room temperature or overnight at 4 °C . Section 3: Primary antibody incubation Carefully remove...wells. Dilute the HRP-conjugated isotype-specific secondary antibody to the desired concentrations in the ...

  10. Immunocytochemistry

    Type
    Protocol
    ...antibody against a target protein and a fluorescent secondary antibody....antibody against a target protein and a fluorescent secondary antibody. This protocol outlines the steps for...serum albumin (BSA) Triton X-100 Primary antibody Secondary antibody Deionized water Microscope slide Anti-fade...incubation times, and recommended compatible reagents. Secondary antibodies must match the host species of the ...primary antibody. For example, use an anti-mouse secondary antibody for primary antibodies raised in a mouse...solution into 5 mL PBS. Protect from light. Procedure Section 1: Seeding cells Place a sterile poly-D-lysine ...to grow to the desired density before labeling. Section 2: Fixing and permeabilizing cells Gently aspirate...

  11. Protocol - How to Streak a Plate

    Type
    Protocol
    ...horizontal lines in another section of the plate, and then diagonal lines in another section of the plate. Make...the first) in each new section crosses at least one line of the previous section so that it will contain...through streak #1 and spread the bacteria over a second section of the plate, to create streak #2. Using a ...the bacteria. Gently spread the bacteria over a section of the plate, as shown in the diagram above, to...streak #2 and spread the bacteria over the last section of the plate, to create streak #3. Incubate plate...

  12. Pipetting Protocol

    Type
    Protocol
    ...read differently on each type of pipette. This section will help you understand how to read each display...065. P200: The first digit is hundreds of µL, the second is tens, and the third is ones. Therefore 100 µL...as 095. P20: The first digit is tens of µL, the second is ones, and the third (red) digit is tenths. Therefore... as 022. P2: The first digit is ones of µL, the second red digit is tenths, and the third (red) digit ...decimal place. For the P20, this represents the second decimal place, and for the P2 this represents the...for an accurate measurement in the next step. The second stop is meant to expel excess liquid from the tip...slowly press down on the plunger all the way to the second stop to release the liquid. Pro-Tip Dispense until...

  13. Kit Free RNA Extraction

    Type
    Protocol
    ...provides two options: using Solution D (see reagent section for recipe) or using an all-in-one acid guanidinium..., make sure to prepare solution D (see reagent section above for the recipe). If using TRIzol®, jump down... down to the Option #2 - TRIzol® Protocol section below. Homogenize or lyse tissues or cells in Solution...49:1) and then shake vigorously by hand for 10 seconds. Incubate sample(s) for 15 minutes on ice and centrifuge...in 0.5–1 ml of 75% Ethanol and vortex for a few seconds. Centrifuge for 5 minutes at 10,000 x g at 4 °C... TRIzol® used. Shake vigorously by hand for 10 seconds. Incubate the sample(s) for 2-3 minutes on ice ...

  14. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Aliquot and freeze upright at -20 °C. Procedure Section 1: Seeding cells The day prior to transfecting,...Pro-Tip Do not use cells that are over 30 passages. Section 2: Transfection Check the cell density and viability... s to mix. Add 450 µL of 1 mg/mL PEI-MAX to the second tube of 6 mL BCD TFX (for a 1 mg/mL stock solution...times to mix. Return the flask to the incubator. Section 3: BCD Feed and valproic acid supplementation During...Thursday (168 h post-transfection): Harvest antibody. Section 4: Harvest antibody At 168 hours (1 week) post-...

  15. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ... 30 seconds at 94°C: Continued denaturation of double stranded DNA. Anneal primers for 30 seconds at 55...minutes at 94°C. Denature for 30 seconds at 94°C. Anneal primers for 30 seconds at 55°C (or 5°C below Tm). ...45µl volume of master mix to add to each PCR tube. Secure the tops to the PCR tubes, gently tap each tube...

  16. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ... by the Gradinaru lab. For simplicity, in each section we’ll list the Gradinaru capsids first, before ... levels to AAV9 (Stanton et al., 2023). PAL2, a second-generation PAL capsid, shows enhanced transduction...

  17. Antibody Guide

    Type
    Guide
    ...similar factors. Secondary antibodies Secondary antibodies, often called secondaries, are usually much...and a conjugated secondary antibody specific to your primary antibody. Multiple secondary antibodies will...if your fluorophore is bound to a secondary antibody, the secondary antibodies must be able to distinguish...Therefore, secondary antibodies are species specific and are often referred to by [secondary source] [primary...[primary source secondary recognizes], e.g. , “goat anti-rabbit.” Choose a secondary that recognizes your...applications, and is covered in the following Isotypes section. The variable region of the antibody is the antigen...the amplification of signals by having multiple secondary antibodies binding to a primary antibody which...

  18. Plasmids 101 eBook 4th Edition

    Type
    Blog Post
    ...edition has been fully updated with twelve new sections, including our popular "Dimers and Multimers" ...

  20. Pouring LB Agar Plates

    Type
    Protocol
    ...prepare your plate pouring station: Find an empty section of lab bench with a working flame. Spray down the... containing the appropriate antibiotics, and a section for active pouring. Prepare your antibiotics. Prior... know to be resistant to the antibiotic. On the second plate, streak out a strain that’s not resistant...temperature and check for growth. See our sample data section below for positive and negative test results. Sample...
Showing: 261 - 280 of 545 results