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We narrowed to 69 results for: addgene/1000

Showing: 21 - 40 of 69 results

  1. 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 ...

  2. 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...

  3. 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...

  4. 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 ...

  5. 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...

  6. 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 ...

  7. 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...

  8. 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....

  9. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you...proteins that result in a diverse set of colors. Addgene has assembled a collection of empty plasmid backbones...exctinction coefficient and quantum yield divided by 1000. pKa is the pH at which fluorescence intensity is...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource! Jump to Top...

  10. Promoters

    Type
    Guide
    ...Addgene's molecular biology reference material covering promoters, transcription and the transcription...interest is expressed. Summary Promoters are about 100-1000 base pairs long and are adjacent and typically upstream...small RNAs, such as shRNAs and gRNAs. Resources Addgene's blog , including our popular Plasmids 101 series...

  11. Antibody Guide

    Type
    Collection
    ...Science Guides Antibody Guide Antibody Guide View Addgene's Antibody Collection Jump To: Antibody Guide Production...applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process may...intensive. Ready to put your new knowledge to use? Addgene now makes ready-to-use recombinant antibodies made...

  12. 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...

  13. 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...

  14. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...plasmids with Addgene. Additional control plasmids for the TKO library will be available at Addgene soon. Hart...cellular compartments (cytosol, Addgene catalog #71245; mitochondria, Addgene catalog #71247) Rosella provides...read? Listen to our hot plasmids segment on the Addgene Podcast. View Plasmids from:  March (Q1) | June...sequencing. The STARR-seq backbone is available from Addgene in both human and fly versions. Arnold, et al....alongside a growing number of CRISPR libraries from Addgene’s repository that can be used to perform screens...PMID: 24395366 Oxford Drosophila CRISPR library Addgene has had the privilege of distributing several human...promoter and Cas9 from the actin 5C promoter. Addgene’s fly library web page offers more details and protocols...

  15. 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...

  16. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    ...antibodies, plasmids (like your Addgene plasmid #), and other reagents. F1000Research, among many other journals...Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place in the world...results more valuable. Some journals, including F1000Research, also ask for the raw data behind the figures...several rounds of revision before acceptance. F1000Research uses a novel form of peer review that occurs... Maaike Pols is a Developmental Editor for F1000Research, responsible for assessing the scientific content...

  17. Molecular Biology Reference

    Type
    Guide
    ...videos for specific techniques and tips! Addgene Protocols Addgene Video Library...powder can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots...Addgene's molecular biology reference guide, including information on molecular genetics, plasmids, sequencing... plasmid. For commonly used primers check out Addgene's sequencing primer list. Working with Plasmids ...empty plasmid backbone for your experiment, see Addgene's empty backbone page for more information. Expression...genome. For more information on CRISPR check out Addgene’s CRISPR guide . Reporter Plasmids - Used for studying...infect your target cells at a high efficiency. Addgene's expanding viral service offers select ready-made...

  18. 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...

  19. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...Heckler. Additional Resources on the Addgene Blog Get Addgene's Take Aways from the FoR 2015 Symposium...sector throughout their careers (McDowell et al, F1000Research, 2014; Dolan, F100Research, 2016). These events...Research: a perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...from the Future of Research Chicago Symposium F1000Research, 2016. Kyle T. Dolan, Joseph F. Pierre and Erin...

  20. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...101: Validation Resources on addgene.org Addgene's Antibody Guide Addgene's Antibody Protocols...10.1038/s41598-019-55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies...and flow cytometry. F1000Research, 13, 817. https://doi.org/10.12688/f1000research.154034.1 Laflamme, ...synaptotagmin 1 page. The data produced is available on the F1000 gateway and Zenodo community.  Knockdown approach...
Showing: 21 - 40 of 69 results