d pbs  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 86
    Name:
    PBS Phosphate Buffered Saline Tablets
    Description:
    Phosphate Buffered Saline PBS is a balanced salt solution used for a variety of applications including reagent preparation diluting cells for flow cytometry and as a cell culture reagent These PBS tablets are not intended for cell culture as they are not tested for sterility We offer a variety of Gibco PBS formulations for cell culture applications Find the right cell culture product using the media selector tool
    Catalog Number:
    003002
    Price:
    None
    Category:
    Lab Reagents and Chemicals
    Applications:
    Cell Analysis|Cellular Imaging|IHC Counterstaining & Mounting|IHC Epitope Retrieval, Blocking & Quenching|IHC Sample Preparation|IHC Staining & Detection|Immunocytochemistry (ICC)|Immunofluorescence (IF)|Immunohistochemistry (IHC)
    		Buy from Supplier

    Structured Review

    Thermo Fisher d pbs
    Phosphate Buffered Saline PBS is a balanced salt solution used for a variety of applications including reagent preparation diluting cells for flow cytometry and as a cell culture reagent These PBS tablets are not intended for cell culture as they are not tested for sterility We offer a variety of Gibco PBS formulations for cell culture applications Find the right cell culture product using the media selector tool
    https://www.bioz.com/result/d pbs/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    d pbs - by Bioz Stars, 2021-03
    86/100 stars

    Images

    Related Articles

    Concentration Assay:

    Article Title: A nanoparticle-based approach to improve the outcome of cancer active immunotherapy with lipopolysaccharides
    Article Snippet: .. Investigation of the 24 h post-injection systemic TNF-α and LPS concentration Serum concentrations of TNF-α and LPS were determined 24 h after the peritumoral injection of the first dose of PBS (control), LPS or LPS-NP at high concentrations (1000 µg/mL) using ELISA (eBioscience) and LAL chromogenic endotoxin quantification assay, respectively. .. Microscopic evaluation of the tumor cross-sections For tracking the LPS penetration (both free and nanoparticle-bound) into the tumor after injection, FITC-labeled LPS was utilized in the preparation of both the solution and nanoparticles, which were injected in three corners around the tumor.

    Injection:

    Article Title: A nanoparticle-based approach to improve the outcome of cancer active immunotherapy with lipopolysaccharides
    Article Snippet: .. Investigation of the 24 h post-injection systemic TNF-α and LPS concentration Serum concentrations of TNF-α and LPS were determined 24 h after the peritumoral injection of the first dose of PBS (control), LPS or LPS-NP at high concentrations (1000 µg/mL) using ELISA (eBioscience) and LAL chromogenic endotoxin quantification assay, respectively. .. Microscopic evaluation of the tumor cross-sections For tracking the LPS penetration (both free and nanoparticle-bound) into the tumor after injection, FITC-labeled LPS was utilized in the preparation of both the solution and nanoparticles, which were injected in three corners around the tumor.

    Enzyme-linked Immunosorbent Assay:

    Article Title: A nanoparticle-based approach to improve the outcome of cancer active immunotherapy with lipopolysaccharides
    Article Snippet: .. Investigation of the 24 h post-injection systemic TNF-α and LPS concentration Serum concentrations of TNF-α and LPS were determined 24 h after the peritumoral injection of the first dose of PBS (control), LPS or LPS-NP at high concentrations (1000 µg/mL) using ELISA (eBioscience) and LAL chromogenic endotoxin quantification assay, respectively. .. Microscopic evaluation of the tumor cross-sections For tracking the LPS penetration (both free and nanoparticle-bound) into the tumor after injection, FITC-labeled LPS was utilized in the preparation of both the solution and nanoparticles, which were injected in three corners around the tumor.

    Incubation:

    Article Title: Potent mouse monoclonal antibodies that block SARS-CoV-2 infection
    Article Snippet: Cells were blocked by 1% non-fat skim milk in PBS-T for 10 min, then incubated with 0.5 µg/mL antibody for 1 h at room temperature. .. After three times wash in PBS-T, cells were incubated in 1:500 diluted Alexa Fluor 488 conjugated goat anti-mouse IgG secondary antibody (Thermo Fisher) and 1 µg/mL DAPI solution for 30 min at room temperature. .. The cover glasses were mounted with Prolong Glass Antifade Mountant (Thermo Fisher) overnight at room temperature before observing.

    Article Title: A purified, fermented, extract of Triticum aestivum has lymphomacidal activity mediated via natural killer cell activation
    Article Snippet: .. To assess caspase activity, cells were incubated with FWGP or PBS control as above and stained with a Vybrant FAM Poly Caspases Assay Kit (Molecular Probes) according to the manufacturer’s instructions. .. Briefly, 300 μl of cell suspension (1 x 106 cells/ml) were incubated with VAD-FMK FLICA reagent and Hoechst 33342 for the detection of activated caspases 1, 2, 4, 5, 6, 8 and 9, washed and analyzed by flow cytometry as above.

    Modification:

    Article Title: Wnt Signaling Induces Epithelial-Mesenchymal Transition with Proliferation in ARPE-19 Cells upon Loss of Contact Inhibition
    Article Snippet: These findings highlight the importance of the Wnt signaling activated by EGF+FGF-2 to unlock contact inhibition in order to initiate an early phase of EMT with proliferation, and establish a framework for future dissection of such pathogenic EMT in proliferative vitreoretinopathy. .. Antibodies and Reagents Dulbecco’s modified Eagle’s medium (DMEM), Ham’s/F12 medium, human epidermal growth factor (EGF), HEPES buffer, phosphate-buffered saline (PBS), amphotericin B, gentamicin, fetal bovine serum (FBS), and Alexa fluor-conjugated secondary IgG were purchased from Invitrogen (Carlsbad, CA). .. FGF-2, TGF-β1, bovine serum albumin, paraformaldehyde, methanol, Triton X-100, XAV939 (tankyrase I inhibitor), and Hoechst 33342 dye were purchased from Sigma (St. Louis, MO).

    Cell Culture:

    Article Title: Renal-targeted delivery of triptolide by entrapment in pegylated TRX-20-modified liposomes
    Article Snippet: .. After enzymatic digestions with collagenase IV (0.1% w/v) at 37°C in PBS solution for 20–45 min, the MC suspensions were obtained and cultured in RPMI 1640 medium containing 20% heat-inactivated fetal bovine serum, 2 μg/mL insulin, 300 μg/L transferrin, 100 U/mL penicillin, and 100 U/mL streptomycin at 37°C in a humidified 5% (v/v) CO2 incubator (Thermo Scientific, Marietta, OH, USA). ..

    other:

    Article Title: Expansion Sequencing: Spatially Precise In Situ Transcriptomics in Intact Biological Systems
    Article Snippet: The reaction underwent heat inactivation at 75°C for 5 min, and finally washed for 1 hour with 1X PBS.

    Activity Assay:

    Article Title: A purified, fermented, extract of Triticum aestivum has lymphomacidal activity mediated via natural killer cell activation
    Article Snippet: .. To assess caspase activity, cells were incubated with FWGP or PBS control as above and stained with a Vybrant FAM Poly Caspases Assay Kit (Molecular Probes) according to the manufacturer’s instructions. .. Briefly, 300 μl of cell suspension (1 x 106 cells/ml) were incubated with VAD-FMK FLICA reagent and Hoechst 33342 for the detection of activated caspases 1, 2, 4, 5, 6, 8 and 9, washed and analyzed by flow cytometry as above.

    Staining:

    Article Title: A purified, fermented, extract of Triticum aestivum has lymphomacidal activity mediated via natural killer cell activation
    Article Snippet: .. To assess caspase activity, cells were incubated with FWGP or PBS control as above and stained with a Vybrant FAM Poly Caspases Assay Kit (Molecular Probes) according to the manufacturer’s instructions. .. Briefly, 300 μl of cell suspension (1 x 106 cells/ml) were incubated with VAD-FMK FLICA reagent and Hoechst 33342 for the detection of activated caspases 1, 2, 4, 5, 6, 8 and 9, washed and analyzed by flow cytometry as above.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • d pbs  (Thermo Fisher)
    97
    Thermo Fisher d pbs
    Over-the-counter (OTC) lubricants reduce epithelial cell integrity. Caco-2 cell monolayers were incubated with each lubricant (triplicates of neat samples) and the TEER measured 2, 4, and 6 h later. <t>Carraguard</t> and Gynol II were included as controls. Data are shown as Ω×cm 2 (mean±SD of the triplicates) averaging the data for all of the lubricants versus the Gynol II, Carraguard, and <t>D-PBS</t> controls.
    D Pbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/d pbs/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    d pbs - by Bioz Stars, 2021-03
    97/100 stars
    		  Buy from Supplier

    Image Search Results


    Over-the-counter (OTC) lubricants reduce epithelial cell integrity. Caco-2 cell monolayers were incubated with each lubricant (triplicates of neat samples) and the TEER measured 2, 4, and 6 h later. Carraguard and Gynol II were included as controls. Data are shown as Ω×cm 2 (mean±SD of the triplicates) averaging the data for all of the lubricants versus the Gynol II, Carraguard, and D-PBS controls.

    Journal: AIDS Research and Human Retroviruses

    Article Title: Identification of Personal Lubricants That Can Cause Rectal Epithelial Cell Damage and Enhance HIV Type 1 Replication in Vitro

    doi: 10.1089/aid.2010.0252

    Figure Lengend Snippet: Over-the-counter (OTC) lubricants reduce epithelial cell integrity. Caco-2 cell monolayers were incubated with each lubricant (triplicates of neat samples) and the TEER measured 2, 4, and 6 h later. Carraguard and Gynol II were included as controls. Data are shown as Ω×cm 2 (mean±SD of the triplicates) averaging the data for all of the lubricants versus the Gynol II, Carraguard, and D-PBS controls.

    Article Snippet: Additionally we used Carraguard (Population Council, New York, NY), D-PBS (Invitrogen, Grand Island, NY), and Gynol II Vaginal Contraceptive Jelly (Caldwell Consumer Health, LLC, Parsippany, NJ).

    Techniques: Incubation

    Zinc acetate in HEC does not prevent vaginal or rectal HSV-2 infection in mice. Medroxyprogesterone acetate-treated (vaginal; A) or untreated (rectal; B) mice had the indicated gels (versus D-PBS or the zinc acetate solution) applied vaginally or rectally 10 min before being challenged with 10 6 PFU (via the respective routes). Survival was monitored for up to 20 days, and the survival curves are shown for each condition ( n = 15 to 20 for each treatment group).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Zinc Acetate/Carrageenan Gels Exhibit Potent Activity In Vivo against High-Dose Herpes Simplex Virus 2 Vaginal and Rectal Challenge

    doi: 10.1128/AAC.05461-11

    Figure Lengend Snippet: Zinc acetate in HEC does not prevent vaginal or rectal HSV-2 infection in mice. Medroxyprogesterone acetate-treated (vaginal; A) or untreated (rectal; B) mice had the indicated gels (versus D-PBS or the zinc acetate solution) applied vaginally or rectally 10 min before being challenged with 10 6 PFU (via the respective routes). Survival was monitored for up to 20 days, and the survival curves are shown for each condition ( n = 15 to 20 for each treatment group).

    Article Snippet: Mice received subcutaneously 100 μl of medroxyprogesterone acetate (Depo-Provera; Upjohn, Kalamazoo, MI) at 25 mg/ml in D-PBS (Invitrogen).

    Techniques: Infection, Mouse Assay

    Repeated treatment with zinc-carrageenan formulations does not increase HSV-2 susceptibility in the mouse model. The indicated gel formulations were delivered intravaginally daily for 7 days to medroxyprogesterone acetate-treated BALB/c mice ( n = 20 to 24 per group). Twelve hours after the last gel was applied, mice were challenged with 2 × 10 3 PFU of HSV-2 strain G. Mice were examined and scored daily for 19 days. Percent survival over time is shown for each treatment group. There was not a significant difference between each individual gel (D, E, or G; P > 0.096) or the combination of the data from those three gels ( P > 0.055) compared to D-PBS or carrageenan gel. A very significant difference ( P = 0.0009 or P = 0.0004) was observed when comparing Gynol versus D-PBS or carrageenan, respectively.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Zinc Acetate/Carrageenan Gels Exhibit Potent Activity In Vivo against High-Dose Herpes Simplex Virus 2 Vaginal and Rectal Challenge

    doi: 10.1128/AAC.05461-11

    Figure Lengend Snippet: Repeated treatment with zinc-carrageenan formulations does not increase HSV-2 susceptibility in the mouse model. The indicated gel formulations were delivered intravaginally daily for 7 days to medroxyprogesterone acetate-treated BALB/c mice ( n = 20 to 24 per group). Twelve hours after the last gel was applied, mice were challenged with 2 × 10 3 PFU of HSV-2 strain G. Mice were examined and scored daily for 19 days. Percent survival over time is shown for each treatment group. There was not a significant difference between each individual gel (D, E, or G; P > 0.096) or the combination of the data from those three gels ( P > 0.055) compared to D-PBS or carrageenan gel. A very significant difference ( P = 0.0009 or P = 0.0004) was observed when comparing Gynol versus D-PBS or carrageenan, respectively.

    Article Snippet: Mice received subcutaneously 100 μl of medroxyprogesterone acetate (Depo-Provera; Upjohn, Kalamazoo, MI) at 25 mg/ml in D-PBS (Invitrogen).

    Techniques: Mouse Assay

    Zinc-carrageenan formulations do not negatively impact the architecture of cervicovaginal and rectal epithelia in vivo . Eight-week-old female BALB/c mice were treated with medroxyprogesterone acetate or fasted and anesthetized before adding D-PBS, carrageenan-based gel, formulation D, formulation G, or Gynol II to evaluate the normal architecture in the cervicovaginal and rectal mucosae. Mice were sacrificed at 1, 6, and 24 h after gel applications, and the entire reproductive or rectal tract was surgically excised, fixed, and embedded in paraffin before preparing tissue sections for morphological analyses using H E staining. Panel A (cervicovaginal mucosa) and panel B (rectal mucosa) represent a sample of the 6 sections of two or three animals that were analyzed per formulation. The original magnification is 20×, and the bar length represents 0.1 mm.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Zinc Acetate/Carrageenan Gels Exhibit Potent Activity In Vivo against High-Dose Herpes Simplex Virus 2 Vaginal and Rectal Challenge

    doi: 10.1128/AAC.05461-11

    Figure Lengend Snippet: Zinc-carrageenan formulations do not negatively impact the architecture of cervicovaginal and rectal epithelia in vivo . Eight-week-old female BALB/c mice were treated with medroxyprogesterone acetate or fasted and anesthetized before adding D-PBS, carrageenan-based gel, formulation D, formulation G, or Gynol II to evaluate the normal architecture in the cervicovaginal and rectal mucosae. Mice were sacrificed at 1, 6, and 24 h after gel applications, and the entire reproductive or rectal tract was surgically excised, fixed, and embedded in paraffin before preparing tissue sections for morphological analyses using H E staining. Panel A (cervicovaginal mucosa) and panel B (rectal mucosa) represent a sample of the 6 sections of two or three animals that were analyzed per formulation. The original magnification is 20×, and the bar length represents 0.1 mm.

    Article Snippet: Mice received subcutaneously 100 μl of medroxyprogesterone acetate (Depo-Provera; Upjohn, Kalamazoo, MI) at 25 mg/ml in D-PBS (Invitrogen).

    Techniques: In Vivo, Mouse Assay, Staining

    N-GFP fusion protein assembles into fluorescent SRS that are internalized by murine macrophage and dendritic cell lines. (A) SDS-PAGE analysis of purified GST-PCT+N-GFP complex. Sample (1) was denatured in Laemmli buffer, run on a 12% polyacrylamide gel and detected with Coomassie brilliant blue staining. (2) protein molecular size standards (kDa). (B) Observation of the different complexes adsorbed on glutathione-Sepharose 4B beads under UV light. (C) Electron micrographs of ring-like structures produced by heterologous expression of N-GFP (left) and N (right) purified by GST-PCT. Bars, 50 nm. (D) N-GFP SRS (thin line) adsorption by RAW and D2SC/1 cell lines after 1 hour incubation at 4°C (controls: PBS in grey or GFP bold line). Green fluorescence associated with living cells was analyzed by flow cytometry (the gate was set up excluding dead cells stained with propidium iodide in FL3). The data (100,000 events) were acquired with a FACScalibur and analyzed with Cell Quest-Pro. (E) Confocal microscopy analysis showing entry of N-GFP SRS (green fluorescence) within cells. Filamentous actin was stained with phalloidin Rhodamin (red). Images of individual cells are either z sections through confocal images taken at sequential focal planes or xy views. Stacks of confocal images were acquired at 0.37 µm intervals (bars 10 µm).

    Journal: PLoS ONE

    Article Title: Sub-Nucleocapsid Nanoparticles: A Nasal Vaccine against Respiratory Syncytial Virus

    doi: 10.1371/journal.pone.0001766

    Figure Lengend Snippet: N-GFP fusion protein assembles into fluorescent SRS that are internalized by murine macrophage and dendritic cell lines. (A) SDS-PAGE analysis of purified GST-PCT+N-GFP complex. Sample (1) was denatured in Laemmli buffer, run on a 12% polyacrylamide gel and detected with Coomassie brilliant blue staining. (2) protein molecular size standards (kDa). (B) Observation of the different complexes adsorbed on glutathione-Sepharose 4B beads under UV light. (C) Electron micrographs of ring-like structures produced by heterologous expression of N-GFP (left) and N (right) purified by GST-PCT. Bars, 50 nm. (D) N-GFP SRS (thin line) adsorption by RAW and D2SC/1 cell lines after 1 hour incubation at 4°C (controls: PBS in grey or GFP bold line). Green fluorescence associated with living cells was analyzed by flow cytometry (the gate was set up excluding dead cells stained with propidium iodide in FL3). The data (100,000 events) were acquired with a FACScalibur and analyzed with Cell Quest-Pro. (E) Confocal microscopy analysis showing entry of N-GFP SRS (green fluorescence) within cells. Filamentous actin was stained with phalloidin Rhodamin (red). Images of individual cells are either z sections through confocal images taken at sequential focal planes or xy views. Stacks of confocal images were acquired at 0.37 µm intervals (bars 10 µm).

    Article Snippet: RAW and D2SC/1 cells were incubated in D-PBS (Gibco) supplemented with 5 mM EDTA, 0.33% lidocaine (Astra Zeneca) for 2 min at 37°C and centrifuged for 7 min at 500 g. The pellet was resuspended in PBS supplemented with 2% FCS.

    Techniques: SDS Page, Purification, Staining, Produced, Expressing, Adsorption, Incubation, Fluorescence, Flow Cytometry, Cytometry, Confocal Microscopy

    Evaluation of exon skipping efficacy 7 days after AAV9-U7-AON-5+6 administration in newborn mice 1-day-old KI mice received PBS or adeno-associated virus serotype 9 (AAV9) encoding U7-AON-5+6 (2 × 10 11 vg) by systemic administration into the temporal vein for 7 days. RT-PCR using primers located in exons 4 and 9 of Mybpc3 or in Gapdh . Determination of the mRNA levels was performed by densitometry (Gene Tool Sofware; Syngene, Cambridge). Total Mybpc3 mRNA level was normalized to Gapdh mRNA level and related to KI-PBS. Var-4 and Mut-1 mRNA levels were expressed as percentage of total Mybpc3 mRNA. Western blot stained with antibodies directed against the MyBP-C motif of cMyBP-C or α-actinin. Total cMyBP-C level was normalized to α-actinin and related to KI-PBS, and Var-4 protein level was expressed as a percentage of total cMyBP-C protein level. Var-4 positive control corresponds to protein extract from HEK293 cells transfected with Var-4 cDNA. Data are expressed as mean ± SEM. * p

    Journal: EMBO Molecular Medicine

    Article Title: Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice

    doi: 10.1002/emmm.201202168

    Figure Lengend Snippet: Evaluation of exon skipping efficacy 7 days after AAV9-U7-AON-5+6 administration in newborn mice 1-day-old KI mice received PBS or adeno-associated virus serotype 9 (AAV9) encoding U7-AON-5+6 (2 × 10 11 vg) by systemic administration into the temporal vein for 7 days. RT-PCR using primers located in exons 4 and 9 of Mybpc3 or in Gapdh . Determination of the mRNA levels was performed by densitometry (Gene Tool Sofware; Syngene, Cambridge). Total Mybpc3 mRNA level was normalized to Gapdh mRNA level and related to KI-PBS. Var-4 and Mut-1 mRNA levels were expressed as percentage of total Mybpc3 mRNA. Western blot stained with antibodies directed against the MyBP-C motif of cMyBP-C or α-actinin. Total cMyBP-C level was normalized to α-actinin and related to KI-PBS, and Var-4 protein level was expressed as a percentage of total cMyBP-C protein level. Var-4 positive control corresponds to protein extract from HEK293 cells transfected with Var-4 cDNA. Data are expressed as mean ± SEM. * p

    Article Snippet: Western blot analysis Crude protein extract was obtained from 50 mg ventricular tissue or from one well (12-well-plate) of cultured NMCMs or HEK293 cells, which was rinsed once with ice-cold 1x D-PBS (Life Technologies).

    Techniques: Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Positive Control, Transfection

    Impact of AAV9-U7-AONs on molecular and functional phenotype in 4-week-old mice KI mice received NaCl, PBS, adeno-associated virus serotype 9 (AAV9) encoding GFP (GFP, 7.6 × 10 10 vg) or encoding U7-AON-5+6 (9.4 × 10 11 vg) by systemic administration into the tail vein. Analyses of ventricular tissue were performed 4 weeks post-injection. RT-PCR analysis using primers located in exons 4 and 9 of Mybpc3 and α-cardiac muscle actin ( Actc1 ). The level of mutant Mybpc3 mRNAs was quantified using the Gene Tool Software (Syngene, Cambridge) and the level of Var-4 mRNA was expressed as percentage of total Mybpc3 mRNAs, and indicated in the figure. Western blot stained with antibodies directed against the N-terminus of cMyBP-C (cMyBP-C), the amino acids produced by the fusion of exon 4 with exon 7 (Var-4) or against α-actinin. The Var-4-cMyBP-C antibody also detects WT, Mut-1 and/or Mut-3 cMyBP-C proteins. As positive controls, protein extracts from either ventricular tissue of a wild-type mouse injected with PBS (WT-NaCl) or from HEK293 cells transfected with a plasmid encoding Var-4 were used (HEK-Var-4). The expected fragments are indicated by arrowheads. Echocardiographic analyses were performed 4 weeks after administration of AAV9 or PBS in KI and/or WT mice. Fractional area shortening (FAS), left ventricular mass-to-body-weight (LVM/BW) ratio are shown. Data are expressed as mean ± SEM. ** p

    Journal: EMBO Molecular Medicine

    Article Title: Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice

    doi: 10.1002/emmm.201202168

    Figure Lengend Snippet: Impact of AAV9-U7-AONs on molecular and functional phenotype in 4-week-old mice KI mice received NaCl, PBS, adeno-associated virus serotype 9 (AAV9) encoding GFP (GFP, 7.6 × 10 10 vg) or encoding U7-AON-5+6 (9.4 × 10 11 vg) by systemic administration into the tail vein. Analyses of ventricular tissue were performed 4 weeks post-injection. RT-PCR analysis using primers located in exons 4 and 9 of Mybpc3 and α-cardiac muscle actin ( Actc1 ). The level of mutant Mybpc3 mRNAs was quantified using the Gene Tool Software (Syngene, Cambridge) and the level of Var-4 mRNA was expressed as percentage of total Mybpc3 mRNAs, and indicated in the figure. Western blot stained with antibodies directed against the N-terminus of cMyBP-C (cMyBP-C), the amino acids produced by the fusion of exon 4 with exon 7 (Var-4) or against α-actinin. The Var-4-cMyBP-C antibody also detects WT, Mut-1 and/or Mut-3 cMyBP-C proteins. As positive controls, protein extracts from either ventricular tissue of a wild-type mouse injected with PBS (WT-NaCl) or from HEK293 cells transfected with a plasmid encoding Var-4 were used (HEK-Var-4). The expected fragments are indicated by arrowheads. Echocardiographic analyses were performed 4 weeks after administration of AAV9 or PBS in KI and/or WT mice. Fractional area shortening (FAS), left ventricular mass-to-body-weight (LVM/BW) ratio are shown. Data are expressed as mean ± SEM. ** p

    Article Snippet: Western blot analysis Crude protein extract was obtained from 50 mg ventricular tissue or from one well (12-well-plate) of cultured NMCMs or HEK293 cells, which was rinsed once with ice-cold 1x D-PBS (Life Technologies).

    Techniques: Functional Assay, Mouse Assay, Injection, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Software, Western Blot, Staining, Produced, Transfection, Plasmid Preparation