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We narrowed to 13 results for: addgene/1000

Showing: 1 - 13 of 13 results

  1. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral Service Introduction This protocol...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller...concentration that was used for the initial selection). Addgene recommends that the use of selective medium throughout...cell lines. When this protocol was performed at Addgene on the A549 cells described in our sample data ...96-well plate, seed one of the corner wells with ~1000 cells. This will help you focus the microscope later... incubating the cells. Pro-Tip Use the well with 1000 cells to focus the microscope, then you can easily... 1 lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors...

  2. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...using ready-to-use bio-safe Coomassie G-250 stain (Addgene uses SimplyBlue SafeStain) and ImageJ software....reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Pipette controller Microcentrifuge...Pipette tips, 200 µL, VWR 76322-150 Pipette tips, 1000 µL, VWR 76322-154 Pipettes, 5 mL, VWR 89130-896 ... may not be functional. This sample would fail Addgene QC and would not be used. Section 3: Image Analysis...11469.296 4655.113 16642.522 16124.409 96.887% Pro-Tip Addgene only uses recombinant antibody preps that have ...

  3. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...mL conical collection tube. Centrifuge column at 1000 x g for 2 min to remove storage solution. Place ...container. Add 5 mL PBS to the column. Centrifuge at 1000 x g for 2 min to remove PBS. Repeat Steps 30–31 ...resin bed to fully absorb the sample. Centrifuge at 1000 x g for 2 min to collect the sample. Discard column... Adjust pH to 7.0 Autoclave or filter sterilize 1000X protease inhibitor cocktail 25 mg leupeptin 50 mg...

  4. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...filter, VWR 431229 0.22 µm PES filtering system, 1000 mL, VWR 431098 0.45 µm PES filtering system, 500... aliquots and store at -20 °C until use. BCD TFX 1000 mL BalanCD HEK293 Media 10 mL 10% Pluronic-F68 40...suggest preparing fresh solutions after one month. 1000X protease inhibitor cocktail 25 mg Leupeptin 50 mg...

  5. Western Blot

    Type
    Protocol
    ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Pipette controller Pipette...your cell pellet but will generally be between 250–1000 µL . The ideal lysis buffer will vary depending ...

  6. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 20–200 µL multichannel...Microcentrifuge tubes, Neptune 3745.X Pipette tips, 1000 µL, VWR 76322-154 Pipette tips, 10 µL, VWR 76322...

  7. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Calculation Size in bp of Addgene plasmid #59462 : 6208 bp Concentration of Addgene plasmid #59462 : 1.07 ...AAV prep using qPCR, SYBR Green Technology, and Addgene’s AAV Titration protocol. Protocols...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...curve: One option is to use plasmid #59462 from Addgene. The values highlighted below in red were calculated...titer >3 x 10 13 GC/mL, use dilutions 5–8 Note: at Addgene, samples typically range from 1 x 10 12 GC/mL to...95 uL 20X 200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution 4 20 uL Dil. 3 80 uL 5X 5000X Dilution ...

  8. Immunocytochemistry

    Type
    Protocol
    ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Reagents 1X PBS Microcentrifuge...

  9. CRISPR Library Amplification

    Type
    Protocol
    ...Molecular Biology Reference How do I process my Addgene pooled library? Introduction Please refer to our...depositor for a specific pooled library available at Addgene. We recommend following the depositor's specific...results. If you obtained the pooled library from Addgene, these protocols can be found on the Pooled Library...MaxiPreps (Qiagen HiSpeed Max, Catalog #12663) Tips (1000 µL, 200 µL, 10 µL) Bacti Cell Spreaders (VWR, 60828...with ice and water to create slurry) 100 mL LB At Addgene we use premixed LB (VWR 101414-072) for convenience...Help Center to get details on how to process your Addgene pooled library... 100 x 100 ÷ 0.1 This number should be at least 1000X greater than number of perturbations in library....

  10. AAV ddPCR Titration

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...calculate the titer for each sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume...control (NTC) should be close to zero (B08). At Addgene, we consider runs with an NTC > 10 to be invalid...

  11. Pipetting Protocol

    Type
    Protocol
    ...middle is hundreds, and the third is tens. Therefore, 1000 µL would read as 100 from top to bottom (as shown...consider using a multichannel pipette. Please watch Addgene’s protocol on multichannel pipetting to learn more...Anatomy of a Pipette This front view photo of a P1000 pipette shows the plunger at the top, which is used...tip off of the pipette. This side view photo of a P1000 pipette shows the volume adjustment ring right below...come in standard sizes: P2, P10, P20, P200, and P1000. The table shows the pipette with the appropriate...This image shows the P2, P10, P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume... 2 to 20 µL P100 10 to 100 µL P200 20 to 200 µL P1000 100 to 1,000 µL Each pipette should be used with...

  12. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors...

  13. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Fluorescence Titration Assay AAV Titration by qPCR Addgene Viral Service Lentivirus Guide Virus Blog Posts...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette...
Showing: 1 - 13 of 13 results