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  1. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...You’ve gotten the plasmid encoding your protein of interest from Addgene, transfected it into your target... keen to study is expressing in your cells? Immunoblotting or simply the western blot, or western, is ...Burnett et al., 1981).  Introduction to western blotting  In a western, proteins are: (1) separated by ...proteins to migrate toward the positive charge at the bottom of the gel and separate by size. Smaller proteins...increasing range of acrylamide content from top to bottom allowing a mix of proteins with a broad size range...gel and membrane are “sandwiched” together with blotting paper and an electric current is applied causing...this method.  Finally, commercially available dry blotting systems efficiently transfer a range of protein...

  2. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    ...the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University ...place?" Many thanks to our guest, Darrell Kotton. Dr. Kotton is the David C. Seldin Professor in the Department...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School... Chapter 5: Research currently under way in the Kotton Lab. One of Darrell's favorite projects - differentiating... as an NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the ...Plasmids from the Kotton Lab Learn more about Stem Cells ... darrell kotton...

  4. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...round-bottom plates using sorter or by using limiting dilution at 30 cells per 96-well round-bottom plate...protospacer sequence and PAM happen to fall on the bottom (Crick) strand while sgRNA-B’s protospacer sequence...protospacer sequence/PAM relative to the top or bottom strand. Determine the reverse complement of each...either grow as individual cells in 96-well flat bottom plate or in a 10 cm dish at low concentration so...derived clones can be picked and moved to a flat bottom 96-well plate. Allow the bulk cells to incubate...Plate 100 μl each into two separate 96-well flat-bottom plates. Keep one plate to allow for clones to grow... Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nature Protocols . 2013. PubMed . Characterization...

  5. Penn Vector Core Partnership with Addgene

    Type
    Collection
    ...(H134R)-mCherry Scott Sternson AV-9-18917P 18917-AAV9 AAV-FLEX-rev-ChR2-tdtomato Scott Sternson AV-5-PV2432...18917-AAV1 AAV-FLEX-rev-ChR2-tdtomato Optogenetics Scott Sternson AV-1-20071P 20071-AAV1 pACAGW-ChR2-Venus-AAV...18916P 18916-AAV1 AAV-FLEX-rev-ChR2(H134R)-mCherry Scott Sternson AV-1-20949P 20949-AAV1 pAAV-double floxed-eNpHR-EYFP-WPRE-pA...10-18917P 18917-AAV1 AAV-FLEX-rev-ChR2-tdtomato Scott Sternson AV-10-PV1963 105542-AAV1 pENN.AAV.CB7.CI.eGFP.WPRE.rBG...-5-18917P 18917-AAV5 AAV-FLEX-rev-ChR2-tdtomato Scott Sternson AV-5-20071P 20071-AAV5 pACAGW-ChR2-Venus-AAV...

  6. Zhang Lab CRISPR Page

    Type
    Collection
    ...specificity of RNA-guided Cas9 nucleases. Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala ...Lin CY, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F. Cell . ...CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nat Protoc . 2013 Nov;8(11):2281-308...cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench...Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O,...

  7. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...alpha ALS Kenneth Scott 116365 pHAGE-ERBB4-S774G ERBB4 EF-1 alpha ALS Kenneth Scott 116736 pHAGE-ERBB4...pBMN-mCherry-Parkin PRKN mCherry Parkinson's Richard Youle 60057 pOTTC293 - pAAV EF1a V5-synuclein (WT) SNCA V5 EF-1 alpha...pHAGE-ERBB4 ERBB4 EF-1 alpha ALS Kenneth Scott 117701 pmSod1-WT-N'His SOD1 His T7 ALS Elizabeth Fisher 117702 pmSod1...CMV-CRY2oligo-mcherry-ANXA11 ANXA11 mCherry CMV ALS Jennifer Lippincott-Schwartz 164204 CMV-CRY2oligo-mcherry-ANXA11(...CMV-CRY2oligo-mcherry-ANXA11(D40G) ANXA11 mCherry CMV ALS Jennifer Lippincott-Schwartz 164205 CMV-CRY2oligo-mcherry-ANXA11(R235Q)...R235Q) ANXA11 mCherry CMV ALS Jennifer Lippincott-Schwartz 164206 CMV-CRY2oligo-mcherry-ANXA11(R346C) ANXA11...ANXA11 mCherry CMV ALS Jennifer Lippincott-Schwartz 164207 CMV-CRY2oligo-mcherry-ANXA11(1-185aa) ANXA11 ...

  8. CRISPR Guide

    Type
    Collection
    ...Harrington, L. B., Doxzen, K. W., Ma, E., Liu, J., Knott, G. J., Edraki, A., Garcia, B., Amrani, N., Chen...Communications , 15 (1), 3663. PMID: 38688943 Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann,...Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott, D. A., & Zhang, F. (2013). Genome engineering using...PMID: 24157548 Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche,...26030444 Slaymaker, I. M., Gao, L., Zetsche, B., Scott, D. A., Yan, W. X., & Zhang, F. (2016). Rationally...Shalem, O., Sanjana, N. E., Hartenian, E., Shi, X., Scott, D. A., Mikkelsen, T. S., Heckl, D., Ebert, B. L... Naito, Y., Nakada, S., Yamamoto, T., Sano, S., Hotta, A., Takeda, J., & Mashimo, T. (2019). CRISPR-Cas3...

  9. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...1829-1838. PMID: 27984730 Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O,...26355004 Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench... PMID: 24336571 Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. 2016. Rationally engineered...Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F. 2017. ...

  10. p53 Pathway

    Type
    Collection
    ...Targets Ther. 2013 Dec 19;7:57-68. doi: 10.2147/OTT.S53876. PubMed PMID: 24379683 . When mutants gain new...powers: news from the mutant p53 field. Brosh R, Rotter V. Nat Rev Cancer. 2009 Oct;9(10):701-13. PubMed...p53 gain-of-function in cancer. Oren, M. and V. Rotter. Cold Spring Harbor Perspectives in Biology. 2010...

  11. Brain Initiative Collection

    Type
    Collection
    ...plasmid Scott Sternson 119742-AAV5 AAV SYN PSAM4 GlyR IRES EGFP Chemogenetics expression plasmid Scott Sternson... Chemogenetics Cre-dependent expression plasmid Scott Sternson 119741-AAV9 AAV SYN flex PSAM4 GlyR IRES...GlyR IRES EGFP Chemogenetics expression plasmid Scott Sternson 123308-AAV9 pAAV-hSyn-GRAB_NE1m Expresses...

  12. Bacterial Expression Systems

    Type
    Collection
    ...imaging, epitope tags like His for easy western blotting or staining, and RNA transcript tags to monitor...without SUMO or TEV cleavage sites to remove tags Scott Gradia Arabinose inducible vectors that come with...without SUMO or TEV cleavage sites to remove tags. Scott Gradia Lactose/IPTG inducible vectors that come ...

  13. CRISPR Guide

    Type
    Guide
    ...Harrington, L. B., Doxzen, K. W., Ma, E., Liu, J., Knott, G. J., Edraki, A., Garcia, B., Amrani, N., Chen...Communications , 15 (1), 3663. PMID: 38688943 Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann,...Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott, D. A., & Zhang, F. (2013). Genome engineering using...PMID: 24157548 Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche,...26030444 Slaymaker, I. M., Gao, L., Zetsche, B., Scott, D. A., Yan, W. X., & Zhang, F. (2016). Rationally...Shalem, O., Sanjana, N. E., Hartenian, E., Shi, X., Scott, D. A., Mikkelsen, T. S., Heckl, D., Ebert, B. L... Naito, Y., Nakada, S., Yamamoto, T., Sano, S., Hotta, A., Takeda, J., & Mashimo, T. (2019). CRISPR-Cas3...

  14. Cre-lox system

    Type
    Collection
    ...Cre and PuroR coexpression EF-1 alpha Lentiviral Kotton 30524 pCSHSP:Cre heat shock inducible Cre Xenopus...EF1aL-Cre-IRES-NeoR-W Cre EF-1 alpha Lentiviral Kotton 128166 pCAGGS-mTagBFP2-T2A-sfGFP-iCre-ERT2 sfGFP-iCre-ERT2...Bacterial Tolonen 137051 pGL2399 DiCre Leishmania Mottram 137859 pCAGGS-mTagBFP2-T2A-iCreERT2 (PAA) iCre-...

  15. CRISPR References and Information

    Type
    Collection
    ...laevis and X. tropicalis ), and zebrafish. Off-Spotter: tool for CRISPR/Cas design Program for designing... Mammalian Cell Lines pSpCas9(BB) (pX330) Link Parrott NEBuilder Assembly of CRISPR vectors using ssDNA...

  16. Deisseroth INTRSECT Collection

    Type
    Collection
    ...Ramakrishnan C, Kozicz T, Deisseroth K, Thiele TE, McElligott ZA, Holmes A, Heisler LK, Kash TL. 2016. Serotonin...Link opens in a new window) Tovote P, Esposito MS, Botta P, Chaudun F, Fadok JP, Markovic M, Wolff SB, Ramakrishnan...

  17. Luciferase Plasmid Collection

    Type
    Collection
    ...expression of firefly luciferase and gene of interest Scott Lowe 18782 MSCV Luciferase PGK-hygro Firefly SV40...SV40 Retroviral expression of firefly luciferase Scott Lowe 14080 pAKlux2 Lux Operon Bacterial expression...

  18. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...60601 Mammalian BamHI/EcoRI none S. pyogenes Hygro Hotta pUC-H1-gRNA 61089 Mammalian BsaI none S. pyogenes... Shuttle 47024 Plant inFusion none S. pyogenes Parrott pCBC-DT1T1 50590 Plant BsaI none S. pyogenes Chen...

  19. Evolution of Lab Techniques

    Type
    Blog Post
    ...evolution. Southern blotting and quantitative real-time PCR Southern blotting, named for its inventor...southern blotting can be used to identify the presence of that mutated gene. In southern blotting, DNA is... look at three historical techniques: southern blotting, restriction mapping, and sequencing gels, as ...signal in comparison to a control sample. Southern blotting applications range from confirming the results... clinical diagnostics. One example of southern blotting being used to identify a gene mutation is the ...test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction digest analysis...When a restriction digest with MstII and southern blotting are conducted on a sample from an individual without...

  20. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...in yeast. Autophagy. 2008 Feb;4(2):205-13. Mark Prescott pH Synaptic vesicle pH sensors for imaging exocytosis...Proc Natl Acad Sci U S A. 2019 Aug 1. Alexander Gottschalk Voltage Near-infrared fluorescent voltage indicator...
Showing: 1 - 20 of 287 results