avidin biotin kit  (Thermo Fisher)


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  • 98
    Name:
    Avidin Biotin Blocking Kit
    Description:
    This kit is used to block endogenous biotin or biotin binding activity in tissue sections The kit contains 18 mL each of Avidin and Biotin solutions
    Catalog Number:
    004303
    Price:
    None
    Applications:
    Cell Analysis|Cellular Imaging|IHC Epitope Retrieval, Blocking & Quenching|Immunohistochemistry (IHC)
    Category:
    Lab Reagents and Chemicals
    Buy from Supplier


    Structured Review

    Thermo Fisher avidin biotin kit
    This kit is used to block endogenous biotin or biotin binding activity in tissue sections The kit contains 18 mL each of Avidin and Biotin solutions
    https://www.bioz.com/result/avidin biotin kit/product/Thermo Fisher
    Average 98 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    avidin biotin kit - by Bioz Stars, 2020-09
    98/100 stars

    Images

    Related Articles

    Immunohistochemistry:

    Article Title: Elevated FGF21 secretion, PGC-1α and ketogenic enzyme expression are hallmarks of iron–sulfur cluster depletion in human skeletal muscle
    Article Snippet: .. For PGC-1α immunohistochemistry, tissue sections were treated with 0.1% H2 O2 in PBS for 10 min to block endogenous peroxidase activity, and endogenous biotin was later blocked following the manufacturer's protocol (Avidin/Biotin Blocking Kit, Invitrogen). .. Samples were blocked and incubated with rabbit anti-PGC-1α antibody (see above) in blocking buffer overnight.

    Amplification:

    Article Title: Practical techniques for detection of Toll-like Receptor-4 (TLR4) in the human Intestine
    Article Snippet: .. Avidin/Biotin blocking kit (Zymed, Carlsbad, CA) stored at 4°C. (see Note 1) Tyramide Signal Amplification (TSA) Biotin System (Perkin Elmer/ NEN Life Science, Boston MA). ..

    Pyrolysis Gas Chromatography:

    Article Title: Elevated FGF21 secretion, PGC-1α and ketogenic enzyme expression are hallmarks of iron–sulfur cluster depletion in human skeletal muscle
    Article Snippet: .. For PGC-1α immunohistochemistry, tissue sections were treated with 0.1% H2 O2 in PBS for 10 min to block endogenous peroxidase activity, and endogenous biotin was later blocked following the manufacturer's protocol (Avidin/Biotin Blocking Kit, Invitrogen). .. Samples were blocked and incubated with rabbit anti-PGC-1α antibody (see above) in blocking buffer overnight.

    Labeling:

    Article Title: ANKS6 is the critical activator of NEK8 kinase in embryonic situs determination and organ patterning
    Article Snippet: .. For anti-ANKS6 IF, a biotin layer was employed to enhance labeling of ANKS6, requiring endogenous biotin to be blocked (Avidin/Biotin blocking kit, Life Technologies) prior to application of the biotin-conjugated secondary antibody (Fab fragment of goat anti-rabbit IgG at 1:250, Jackson Immunochemicals) and streptavidin-Alexa 488 at 1:400. .. Slides were incubated in DAPI and mounted with Prolong Gold (Life Technologies).

    Avidin-Biotin Assay:

    Article Title: Characterisation of 11?-hydroxysteroid dehydrogenase 1 in human orbital adipose tissue: a comparison with subcutaneous and omental fat
    Article Snippet: .. A biotin block was performed on the sections for 10 min using the Zymed avidin/biotin blocking kit (Invitrogen Ltd) before incubating with primary antibody (CD68 clone PG-M1 (Dako, Carpinteria, CA, USA) 1:100 dilution) in 10% (v/v) normal swine serum in TBS buffer for 60 min. .. Negative control sections were incubated in TBS buffer without primary antibody.

    Article Title: Practical techniques for detection of Toll-like Receptor-4 (TLR4) in the human Intestine
    Article Snippet: .. Avidin/Biotin blocking kit (Zymed, Carlsbad, CA) stored at 4°C. (see Note 1) Tyramide Signal Amplification (TSA) Biotin System (Perkin Elmer/ NEN Life Science, Boston MA). ..

    Article Title: Bacterial polyphosphates interfere with the innate host defense to infection
    Article Snippet: .. After treating the surfaces with biotin/streptavidin blocking reagent (Avidin/Biotin Blocking Kit, Thermo Fisher), preformed complexes of 50 µM biotin-labeled L-polyphosphates and Streptavidin-PE or Streptavidin-FITC (BioLegend), generated by 30-min incubation of the reagents on ice, were added to the cells for 1 h at 37 °C. .. LysoTracker Green DND-26 (100 nM, Thermo Fisher Scientific) or CellMask Orange Plasma Membrane Stain (5 µg/ml, Thermo Fisher Scientific) were added gently together with the nuclear dye, Hoechst 33342 (Thermo Fisher Scientific), for another 30 min of incubation.

    Article Title: ANKS6 is the critical activator of NEK8 kinase in embryonic situs determination and organ patterning
    Article Snippet: .. For anti-ANKS6 IF, a biotin layer was employed to enhance labeling of ANKS6, requiring endogenous biotin to be blocked (Avidin/Biotin blocking kit, Life Technologies) prior to application of the biotin-conjugated secondary antibody (Fab fragment of goat anti-rabbit IgG at 1:250, Jackson Immunochemicals) and streptavidin-Alexa 488 at 1:400. .. Slides were incubated in DAPI and mounted with Prolong Gold (Life Technologies).

    Article Title: Elevated FGF21 secretion, PGC-1α and ketogenic enzyme expression are hallmarks of iron–sulfur cluster depletion in human skeletal muscle
    Article Snippet: .. For PGC-1α immunohistochemistry, tissue sections were treated with 0.1% H2 O2 in PBS for 10 min to block endogenous peroxidase activity, and endogenous biotin was later blocked following the manufacturer's protocol (Avidin/Biotin Blocking Kit, Invitrogen). .. Samples were blocked and incubated with rabbit anti-PGC-1α antibody (see above) in blocking buffer overnight.

    Article Title: Targeting the GM-CSF receptor for the treatment of CNS autoimmunity
    Article Snippet: .. Endogenous biotin was blocked with the Avidin/Biotin blocking kit (Invitrogen) when required. .. Non-specific immunoglobulin binding was blocked with serum for 30 min.

    Article Title: Long-term expansion of primary equine keratinocytes that maintain the ability to differentiate into stratified epidermis
    Article Snippet: .. Briefly, slides were treated with 3% hydrogen peroxide and with an avidin/biotin blocking kit (Invitrogen). .. The slides were exposed to 10% normal horse serum and to a primary antibodies for CK14 (Abcam, Cambridge, MA, USA no. ab7800, 1:300) diluted in Tris-buffered saline with 0.5% Tween (TBST) for 1 h at room temperature.

    Article Title: Magnetic Resonance Imaging of Ferumoxide-Labeled Mesenchymal Stem Cells in Cartilage Defects: In Vitro and In Vivo Investigations
    Article Snippet: .. For fluorescent staining, slides were blocked with an avidin-biotin blocking kit (Molecular Probes, Eugene, OR) followed by blocking with 6% normal horse serum. .. Steptavidin-peroxidase conjugate coupled with biotinylated secondary antibody was visualized by Alexa Fluor 594 dye using tyramide amplification technique (TSA HRP-streptavidin kit, Molecular Probes).

    Incubation:

    Article Title: Bacterial polyphosphates interfere with the innate host defense to infection
    Article Snippet: .. After treating the surfaces with biotin/streptavidin blocking reagent (Avidin/Biotin Blocking Kit, Thermo Fisher), preformed complexes of 50 µM biotin-labeled L-polyphosphates and Streptavidin-PE or Streptavidin-FITC (BioLegend), generated by 30-min incubation of the reagents on ice, were added to the cells for 1 h at 37 °C. .. LysoTracker Green DND-26 (100 nM, Thermo Fisher Scientific) or CellMask Orange Plasma Membrane Stain (5 µg/ml, Thermo Fisher Scientific) were added gently together with the nuclear dye, Hoechst 33342 (Thermo Fisher Scientific), for another 30 min of incubation.

    Activity Assay:

    Article Title: Elevated FGF21 secretion, PGC-1α and ketogenic enzyme expression are hallmarks of iron–sulfur cluster depletion in human skeletal muscle
    Article Snippet: .. For PGC-1α immunohistochemistry, tissue sections were treated with 0.1% H2 O2 in PBS for 10 min to block endogenous peroxidase activity, and endogenous biotin was later blocked following the manufacturer's protocol (Avidin/Biotin Blocking Kit, Invitrogen). .. Samples were blocked and incubated with rabbit anti-PGC-1α antibody (see above) in blocking buffer overnight.

    Blocking Assay:

    Article Title: Characterisation of 11?-hydroxysteroid dehydrogenase 1 in human orbital adipose tissue: a comparison with subcutaneous and omental fat
    Article Snippet: .. A biotin block was performed on the sections for 10 min using the Zymed avidin/biotin blocking kit (Invitrogen Ltd) before incubating with primary antibody (CD68 clone PG-M1 (Dako, Carpinteria, CA, USA) 1:100 dilution) in 10% (v/v) normal swine serum in TBS buffer for 60 min. .. Negative control sections were incubated in TBS buffer without primary antibody.

    Article Title: Practical techniques for detection of Toll-like Receptor-4 (TLR4) in the human Intestine
    Article Snippet: .. Avidin/Biotin blocking kit (Zymed, Carlsbad, CA) stored at 4°C. (see Note 1) Tyramide Signal Amplification (TSA) Biotin System (Perkin Elmer/ NEN Life Science, Boston MA). ..

    Article Title: Bacterial polyphosphates interfere with the innate host defense to infection
    Article Snippet: .. After treating the surfaces with biotin/streptavidin blocking reagent (Avidin/Biotin Blocking Kit, Thermo Fisher), preformed complexes of 50 µM biotin-labeled L-polyphosphates and Streptavidin-PE or Streptavidin-FITC (BioLegend), generated by 30-min incubation of the reagents on ice, were added to the cells for 1 h at 37 °C. .. LysoTracker Green DND-26 (100 nM, Thermo Fisher Scientific) or CellMask Orange Plasma Membrane Stain (5 µg/ml, Thermo Fisher Scientific) were added gently together with the nuclear dye, Hoechst 33342 (Thermo Fisher Scientific), for another 30 min of incubation.

    Article Title: ANKS6 is the critical activator of NEK8 kinase in embryonic situs determination and organ patterning
    Article Snippet: .. For anti-ANKS6 IF, a biotin layer was employed to enhance labeling of ANKS6, requiring endogenous biotin to be blocked (Avidin/Biotin blocking kit, Life Technologies) prior to application of the biotin-conjugated secondary antibody (Fab fragment of goat anti-rabbit IgG at 1:250, Jackson Immunochemicals) and streptavidin-Alexa 488 at 1:400. .. Slides were incubated in DAPI and mounted with Prolong Gold (Life Technologies).

    Article Title: Elevated FGF21 secretion, PGC-1α and ketogenic enzyme expression are hallmarks of iron–sulfur cluster depletion in human skeletal muscle
    Article Snippet: .. For PGC-1α immunohistochemistry, tissue sections were treated with 0.1% H2 O2 in PBS for 10 min to block endogenous peroxidase activity, and endogenous biotin was later blocked following the manufacturer's protocol (Avidin/Biotin Blocking Kit, Invitrogen). .. Samples were blocked and incubated with rabbit anti-PGC-1α antibody (see above) in blocking buffer overnight.

    Article Title: Targeting the GM-CSF receptor for the treatment of CNS autoimmunity
    Article Snippet: .. Endogenous biotin was blocked with the Avidin/Biotin blocking kit (Invitrogen) when required. .. Non-specific immunoglobulin binding was blocked with serum for 30 min.

    Article Title: Long-term expansion of primary equine keratinocytes that maintain the ability to differentiate into stratified epidermis
    Article Snippet: .. Briefly, slides were treated with 3% hydrogen peroxide and with an avidin/biotin blocking kit (Invitrogen). .. The slides were exposed to 10% normal horse serum and to a primary antibodies for CK14 (Abcam, Cambridge, MA, USA no. ab7800, 1:300) diluted in Tris-buffered saline with 0.5% Tween (TBST) for 1 h at room temperature.

    Article Title: Magnetic Resonance Imaging of Ferumoxide-Labeled Mesenchymal Stem Cells in Cartilage Defects: In Vitro and In Vivo Investigations
    Article Snippet: .. For fluorescent staining, slides were blocked with an avidin-biotin blocking kit (Molecular Probes, Eugene, OR) followed by blocking with 6% normal horse serum. .. Steptavidin-peroxidase conjugate coupled with biotinylated secondary antibody was visualized by Alexa Fluor 594 dye using tyramide amplification technique (TSA HRP-streptavidin kit, Molecular Probes).

    Staining:

    Article Title: Magnetic Resonance Imaging of Ferumoxide-Labeled Mesenchymal Stem Cells in Cartilage Defects: In Vitro and In Vivo Investigations
    Article Snippet: .. For fluorescent staining, slides were blocked with an avidin-biotin blocking kit (Molecular Probes, Eugene, OR) followed by blocking with 6% normal horse serum. .. Steptavidin-peroxidase conjugate coupled with biotinylated secondary antibody was visualized by Alexa Fluor 594 dye using tyramide amplification technique (TSA HRP-streptavidin kit, Molecular Probes).

    Generated:

    Article Title: Bacterial polyphosphates interfere with the innate host defense to infection
    Article Snippet: .. After treating the surfaces with biotin/streptavidin blocking reagent (Avidin/Biotin Blocking Kit, Thermo Fisher), preformed complexes of 50 µM biotin-labeled L-polyphosphates and Streptavidin-PE or Streptavidin-FITC (BioLegend), generated by 30-min incubation of the reagents on ice, were added to the cells for 1 h at 37 °C. .. LysoTracker Green DND-26 (100 nM, Thermo Fisher Scientific) or CellMask Orange Plasma Membrane Stain (5 µg/ml, Thermo Fisher Scientific) were added gently together with the nuclear dye, Hoechst 33342 (Thermo Fisher Scientific), for another 30 min of incubation.

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  • 99
    Thermo Fisher cell surface protein isolation kit
    <t>Isolation</t> of the <t>cell</t> <t>surface</t> <t>protein</t> fraction and their cellular distribution. Cell surface protein fractions were isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. The cell surface protein was biotinylated and affinity purified with avidin beads. ( A ) Marker proteins in the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and the J1.1 cells. The whole cell, flow through, and affinity purified fractions were subjected to western blotting using anti-HSP40, anti-Calnexin and anti-GAPDH Abs. The 1× and 10× input were resolved on SDS-PAGE. ( B ) Silver staining of the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. Stars indicate protein bands enriched in the cell surface fraction of the stimulated Jurkat cells, as compared with those of the unstimulated Jurkat cells. The 1× and 200× input were resolved on SDS-PAGE. ( C ) LFA-1 expression on the cell surface. The fractions shown in (B) were subjected to western blotting using anti-LFA-1α and anti-β-actin mAbs, and anti-Lck Ab. ( D ) J1.1 cells were prelabeled with CellTracker and were cocultured with unstimulated or stimulated Jurkat cells with cell contact. At 3 days post coculture, the cells were immunostained with anti-HIV-1 gp120 Ab (red) and anti-HLA I mAb or anti-Lck Ab (green). Representative images were shown. Scale bar, 5 µm.
    Cell Surface Protein Isolation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 603 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cell surface protein isolation kit/product/Thermo Fisher
    Average 99 stars, based on 603 article reviews
    Price from $9.99 to $1999.99
    cell surface protein isolation kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    94
    Thermo Fisher pierce pull down biotinylated protein
    LAR structure, sequence alignment and pulldown analysis A: BLAST alignment of the known sequences of mouse, rat and human LAR, PTPσ, PTPδ and PTPµ. The wedge domain of each protein is aligned within the box. B: The tandem intracellular phosphatase domains of human LAR with the previously characterized wedge domain (red) 14 . C-D: Pulldown of recombinant PTPσ with <t>biotinylated</t> ISP or ILP. E-F : Eluted lysate following pulldown was probed with antibodies against either LAR or pan-Nogo receptors. Input is 10% lysate control.
    Pierce Pull Down Biotinylated Protein, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pierce pull down biotinylated protein/product/Thermo Fisher
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    pierce pull down biotinylated protein - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Thermo Fisher annexin v propidium iodide apoptosis assay kit
    Analysis of <t>apoptosis</t> and necrosis by <t>propidium</t> iodide staining and flow cytometry. HSC-T6 cells were treated with the avidin-, neutravidin- and streptavidin-based nanocomplexes (100 nM PCBP2 siRNA) for 24 h and then subjected to the analysis of apoptosis and necrosis. The acquisition data were divided into four quadrants according to the type of fluorescence emitted from the cells: Ql-1 calculates the percent of cells undergoing apoptosis (Annexin V), Q2–1 calculates the percent of cells undergoing late apoptosis or induced necrosis (Annexin V and Propidium Iodide), Q3–1 calculates the percent of cells with no fluorescence and Q4–1 calculates the percent of cells undergoing necrosis induced by the nanocomplex. Results are represented as the mean±SD (n=3).
    Annexin V Propidium Iodide Apoptosis Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/annexin v propidium iodide apoptosis assay kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    annexin v propidium iodide apoptosis assay kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Isolation of the cell surface protein fraction and their cellular distribution. Cell surface protein fractions were isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. The cell surface protein was biotinylated and affinity purified with avidin beads. ( A ) Marker proteins in the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and the J1.1 cells. The whole cell, flow through, and affinity purified fractions were subjected to western blotting using anti-HSP40, anti-Calnexin and anti-GAPDH Abs. The 1× and 10× input were resolved on SDS-PAGE. ( B ) Silver staining of the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. Stars indicate protein bands enriched in the cell surface fraction of the stimulated Jurkat cells, as compared with those of the unstimulated Jurkat cells. The 1× and 200× input were resolved on SDS-PAGE. ( C ) LFA-1 expression on the cell surface. The fractions shown in (B) were subjected to western blotting using anti-LFA-1α and anti-β-actin mAbs, and anti-Lck Ab. ( D ) J1.1 cells were prelabeled with CellTracker and were cocultured with unstimulated or stimulated Jurkat cells with cell contact. At 3 days post coculture, the cells were immunostained with anti-HIV-1 gp120 Ab (red) and anti-HLA I mAb or anti-Lck Ab (green). Representative images were shown. Scale bar, 5 µm.

    Journal: Viruses

    Article Title: HIV Reactivation in Latently Infected Cells with Virological Synapse-Like Cell Contact

    doi: 10.3390/v12040417

    Figure Lengend Snippet: Isolation of the cell surface protein fraction and their cellular distribution. Cell surface protein fractions were isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. The cell surface protein was biotinylated and affinity purified with avidin beads. ( A ) Marker proteins in the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and the J1.1 cells. The whole cell, flow through, and affinity purified fractions were subjected to western blotting using anti-HSP40, anti-Calnexin and anti-GAPDH Abs. The 1× and 10× input were resolved on SDS-PAGE. ( B ) Silver staining of the cell fractions isolated from the unstimulated and stimulated Jurkat cells, and J1.1 cells. Stars indicate protein bands enriched in the cell surface fraction of the stimulated Jurkat cells, as compared with those of the unstimulated Jurkat cells. The 1× and 200× input were resolved on SDS-PAGE. ( C ) LFA-1 expression on the cell surface. The fractions shown in (B) were subjected to western blotting using anti-LFA-1α and anti-β-actin mAbs, and anti-Lck Ab. ( D ) J1.1 cells were prelabeled with CellTracker and were cocultured with unstimulated or stimulated Jurkat cells with cell contact. At 3 days post coculture, the cells were immunostained with anti-HIV-1 gp120 Ab (red) and anti-HLA I mAb or anti-Lck Ab (green). Representative images were shown. Scale bar, 5 µm.

    Article Snippet: Preparation and Analysis of Cell Surface Fractions The isolation of cell surface proteins was performed using Pierce Cell Surface Protein Isolation Kit (Thermo Fisher Scientific).

    Techniques: Isolation, Affinity Purification, Avidin-Biotin Assay, Marker, Western Blot, SDS Page, Silver Staining, Expressing

    STIP1 in ovarian cancer cells is transported to cell membrane for secretion into culture medium. A , after 24-h culture of various ovarian cancer cells, the culture medium supernatant contained STIP1 that was secreted by cultured cells. Absence of GAPDH in the culture media of five cell lines indicates no leakage of intracellular contents into the culture media, suggesting that clearly detected STIP1 in all culture media was from secretion by the cells. In this experiment, however, leakage of intracellular GAPDH was detected in TOV21G and OVCAR3 cells, corresponding to the occurrence of their cell lysis. B , detection of STIP1 on the outer cell membrane of ovarian cancer lines supports its outward transportation. Using the Pierce Cell Surface Protein Isolation kit (Thermo Scientific), we labeled outer membrane proteins on live ovarian cancer cells with biotin, purified them with avidin-agarose beads, and analyzed them with Western blot analysis. C , no trace of HSP90 was identified in the purified cell surface proteins ( left panel ), whereas HSP90 was clearly detected in the total cell lysates ( right panel ). M , molecular mass markers.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Stress-induced Phosphoprotein 1 as a Secreted Biomarker for Human Ovarian Cancer Promotes Cancer Cell Proliferation *

    doi: 10.1074/mcp.M110.000802

    Figure Lengend Snippet: STIP1 in ovarian cancer cells is transported to cell membrane for secretion into culture medium. A , after 24-h culture of various ovarian cancer cells, the culture medium supernatant contained STIP1 that was secreted by cultured cells. Absence of GAPDH in the culture media of five cell lines indicates no leakage of intracellular contents into the culture media, suggesting that clearly detected STIP1 in all culture media was from secretion by the cells. In this experiment, however, leakage of intracellular GAPDH was detected in TOV21G and OVCAR3 cells, corresponding to the occurrence of their cell lysis. B , detection of STIP1 on the outer cell membrane of ovarian cancer lines supports its outward transportation. Using the Pierce Cell Surface Protein Isolation kit (Thermo Scientific), we labeled outer membrane proteins on live ovarian cancer cells with biotin, purified them with avidin-agarose beads, and analyzed them with Western blot analysis. C , no trace of HSP90 was identified in the purified cell surface proteins ( left panel ), whereas HSP90 was clearly detected in the total cell lysates ( right panel ). M , molecular mass markers.

    Article Snippet: Cell surface proteins of ovarian cancer cells were isolated using the Pierce Cell Surface Protein Isolation kit (Thermo Scientific) as described by the manufacturer.

    Techniques: Cell Culture, Lysis, Isolation, Labeling, Purification, Avidin-Biotin Assay, Western Blot

    LAR structure, sequence alignment and pulldown analysis A: BLAST alignment of the known sequences of mouse, rat and human LAR, PTPσ, PTPδ and PTPµ. The wedge domain of each protein is aligned within the box. B: The tandem intracellular phosphatase domains of human LAR with the previously characterized wedge domain (red) 14 . C-D: Pulldown of recombinant PTPσ with biotinylated ISP or ILP. E-F : Eluted lysate following pulldown was probed with antibodies against either LAR or pan-Nogo receptors. Input is 10% lysate control.

    Journal: Nature

    Article Title: Modulation of the proteoglycan receptor PTPσ promotes recovery after spinal cord injury

    doi: 10.1038/nature13974

    Figure Lengend Snippet: LAR structure, sequence alignment and pulldown analysis A: BLAST alignment of the known sequences of mouse, rat and human LAR, PTPσ, PTPδ and PTPµ. The wedge domain of each protein is aligned within the box. B: The tandem intracellular phosphatase domains of human LAR with the previously characterized wedge domain (red) 14 . C-D: Pulldown of recombinant PTPσ with biotinylated ISP or ILP. E-F : Eluted lysate following pulldown was probed with antibodies against either LAR or pan-Nogo receptors. Input is 10% lysate control.

    Article Snippet: Biotinylated Peptide Pulldown For pulldown experiments, we used the Pierce Pull-Down Biotinylated Protein:Protein Interaction Kit (Thermo Scientific 21115).

    Techniques: Sequencing, Recombinant

    Identification and characterization of ISP A : PTPσ structure and wedge domain (red). B : Peptide Sequences. C-F : Pulldown of human, rat and mouse PTPσ with biotinylated ISP. *Nonspecific recognition of PTPδ. G-I : CSPG gradient crossing assay. Dashed Lines=CSPG gradient, scale Bar 50µm, n > 16 gradients/group. J : ISP treatment on PTPσ null neurons (n=12/group). K : Time-lapse imaging of an adult sensory neuron growth cone following 2.5µM ISP treatment ( Supplementary Video 4 ).Time-stamp=minutes. Scale bar 20µM. L : The number of neurons released from a CSPG-rich substrate following agitation (n=28 vehicle/ILP, 16 ISP wells/group). Scale bar 50µm. Error bars=SEM, One way ANOVA, Tukey’s post hoc test, *p

    Journal: Nature

    Article Title: Modulation of the proteoglycan receptor PTPσ promotes recovery after spinal cord injury

    doi: 10.1038/nature13974

    Figure Lengend Snippet: Identification and characterization of ISP A : PTPσ structure and wedge domain (red). B : Peptide Sequences. C-F : Pulldown of human, rat and mouse PTPσ with biotinylated ISP. *Nonspecific recognition of PTPδ. G-I : CSPG gradient crossing assay. Dashed Lines=CSPG gradient, scale Bar 50µm, n > 16 gradients/group. J : ISP treatment on PTPσ null neurons (n=12/group). K : Time-lapse imaging of an adult sensory neuron growth cone following 2.5µM ISP treatment ( Supplementary Video 4 ).Time-stamp=minutes. Scale bar 20µM. L : The number of neurons released from a CSPG-rich substrate following agitation (n=28 vehicle/ILP, 16 ISP wells/group). Scale bar 50µm. Error bars=SEM, One way ANOVA, Tukey’s post hoc test, *p

    Article Snippet: Biotinylated Peptide Pulldown For pulldown experiments, we used the Pierce Pull-Down Biotinylated Protein:Protein Interaction Kit (Thermo Scientific 21115).

    Techniques: Imaging

    Analysis of apoptosis and necrosis by propidium iodide staining and flow cytometry. HSC-T6 cells were treated with the avidin-, neutravidin- and streptavidin-based nanocomplexes (100 nM PCBP2 siRNA) for 24 h and then subjected to the analysis of apoptosis and necrosis. The acquisition data were divided into four quadrants according to the type of fluorescence emitted from the cells: Ql-1 calculates the percent of cells undergoing apoptosis (Annexin V), Q2–1 calculates the percent of cells undergoing late apoptosis or induced necrosis (Annexin V and Propidium Iodide), Q3–1 calculates the percent of cells with no fluorescence and Q4–1 calculates the percent of cells undergoing necrosis induced by the nanocomplex. Results are represented as the mean±SD (n=3).

    Journal: Molecular pharmaceutics

    Article Title: Comparison of avidin, neutravidin, and streptavidin as nanocarriers for efficient siRNA Delivery

    doi: 10.1021/acs.molpharmaceut.6b00933

    Figure Lengend Snippet: Analysis of apoptosis and necrosis by propidium iodide staining and flow cytometry. HSC-T6 cells were treated with the avidin-, neutravidin- and streptavidin-based nanocomplexes (100 nM PCBP2 siRNA) for 24 h and then subjected to the analysis of apoptosis and necrosis. The acquisition data were divided into four quadrants according to the type of fluorescence emitted from the cells: Ql-1 calculates the percent of cells undergoing apoptosis (Annexin V), Q2–1 calculates the percent of cells undergoing late apoptosis or induced necrosis (Annexin V and Propidium Iodide), Q3–1 calculates the percent of cells with no fluorescence and Q4–1 calculates the percent of cells undergoing necrosis induced by the nanocomplex. Results are represented as the mean±SD (n=3).

    Article Snippet: Annexin V–Propidium iodide apoptosis assay kit was obtained from Thermo Fisher Scientific (Grand Island, NY). iTaq™ Universal SYBR® Green One-Step Kit was purchased from Bio-Rad (Hercules, California).

    Techniques: Staining, Flow Cytometry, Cytometry, Avidin-Biotin Assay, Fluorescence