Structured Review

Abcam pbs t
Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in <t>PBS-T,</t> the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).
Pbs T, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 470 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pbs t/product/Abcam
Average 92 stars, based on 470 article reviews
Price from $9.99 to $1999.99
pbs t - by Bioz Stars, 2020-07
92/100 stars

Images

1) Product Images from "Benzamil sensitive ion channels contribute to volume regulation in canine chondrocytes"

Article Title: Benzamil sensitive ion channels contribute to volume regulation in canine chondrocytes

Journal: British Journal of Pharmacology

doi: 10.1111/j.1476-5381.2012.02185.x

Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).
Figure Legend Snippet: Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).

Techniques Used: Immunohistochemistry, Patch Clamp, Expressing, Immunofluorescence, Staining, Concentration Assay, Affinity Purification, Fluorescence, Negative Control, Activity Assay, Mass Spectrometry

Related Articles

Affinity Purification:

Article Title: Benzamil sensitive ion channels contribute to volume regulation in canine chondrocytes
Article Snippet: .. After three washes with PBS-T, the cells were incubated for 2 h with a goat polyclonal secondary antibody to rabbit IgG (Fc-specific, affinity-purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam, Cambridge, MA, USA; ab98462) diluted according to the manufacturer's recommendations (typically diluted 1:200). .. After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence).

Binding Assay:

Article Title: Placental extracellular vesicles express active dipeptidyl peptidase IV; levels are increased in gestational diabetes mellitus
Article Snippet: .. Slides were rinsed with distilled water prior to blocking for non-specific antibody binding using 10% Fetal Calf Serum (FCS) (Sigma Aldrich, Gillingham, UK) in PBS-T (PBS with Tween-20) at R/T for 1 h. Sections were incubated overnight at 4°C with 1% FCS and 0.5 µg/ml anti-CD26 primary antibody (Abcam, Cambridge, UK) or non-immune mouse IgG1 (Biolegend, London, UK) in PBS-T. .. Sections were washed in PBS and incubated with anti-mouse IgG secondary antibody (Life Technologies, Paisley, UK) in a humidifying chamber at R/T for 1 h. Slides were washed with 0.01% PBS-T before being stained with DAB (Vector Laboratories, Peterborough, UK).

Incubation:

Article Title: An in vitro culture model to study the dynamics of colonic microbiota in Syrian golden hamsters and their susceptibility to infection with Clostridium difficile
Article Snippet: .. Hamster sera were serially diluted (fivefold steps) on the plates and incubated at room temperature for 1.5 h. Plates were washed with 300 μl of PBS-T three times and incubated with goat anti-hamster immunoglobulin G horse-radish peroxidase conjugated antibody (Abcam, Cambrige, MA, USA) for 1 h at room temperature. .. Following an additional wash with 300 μl of PBS-T, 50 μl of 3,3′,5,5′-tetramethylbenzidine substrate (Thermo Labsystems, Milford, MA, USA) was added and after 15 min, the reaction was stopped by the addition of 50 μl of 0.4 N sulfuric acid (Fisher Scientific, Pittsburg, PA, USA) and absorbance was measured at 450 nm.

Article Title: mTOR Modulates Methamphetamine-Induced Toxicity through Cell Clearing Systems
Article Snippet: .. Afterwards, cells were incubated overnight at 4°C in 1% BSA in PBS-T containing 1 : 50 anti-LC3 antibody (Abcam) and 1 : 30 anti-P20S (Abcam). .. Finally, cells were incubated for 1 h with fluorophore-conjugated secondary antibodies (1 : 200; goat anti-rabbit Alexa 488 and goat anti-mouse Alexa 594, Molecular Probes, Life Technologies) in 1% BSA in PBS-T at room temperature.

Article Title: Placental extracellular vesicles express active dipeptidyl peptidase IV; levels are increased in gestational diabetes mellitus
Article Snippet: .. Slides were rinsed with distilled water prior to blocking for non-specific antibody binding using 10% Fetal Calf Serum (FCS) (Sigma Aldrich, Gillingham, UK) in PBS-T (PBS with Tween-20) at R/T for 1 h. Sections were incubated overnight at 4°C with 1% FCS and 0.5 µg/ml anti-CD26 primary antibody (Abcam, Cambridge, UK) or non-immune mouse IgG1 (Biolegend, London, UK) in PBS-T. .. Sections were washed in PBS and incubated with anti-mouse IgG secondary antibody (Life Technologies, Paisley, UK) in a humidifying chamber at R/T for 1 h. Slides were washed with 0.01% PBS-T before being stained with DAB (Vector Laboratories, Peterborough, UK).

Article Title: Locked and proteolysis-based transcription activator-like effector (TALE) regulation
Article Snippet: .. Blocked membranes were incubated for 1.5 h with a solution of primary antibodies (rabbit anti-FLAG (Sigma) in 0.2% I-Block, washed four times with PBS-T, incubated with a solution of HRP conjugated secondary antibodies (Goat Anti-Rabbit IgG H & L (Abcam)) and again washed four times with PBS-T. .. Detection was performed with SuperSignal West Pico Chemiluminescent Substrate (Life Technologies) in the G:BOX (Syngene) detector and the figures processed using the ImageJ software.

Article Title: Benzamil sensitive ion channels contribute to volume regulation in canine chondrocytes
Article Snippet: .. After three washes with PBS-T, the cells were incubated for 2 h with a goat polyclonal secondary antibody to rabbit IgG (Fc-specific, affinity-purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam, Cambridge, MA, USA; ab98462) diluted according to the manufacturer's recommendations (typically diluted 1:200). .. After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence).

Labeling:

Article Title: Fingolimod modulates microglial activation to augment markers of remyelination
Article Snippet: .. After washing three times for 1 hour under constant inverting rotation with PBS-T, aggregates were labeled with a second primary antibody overnight (rabbit anti-MBP, AbCam) using the same method. .. Following a further wash, relevant fluorochrome-conjugated secondary antibodies were applied and aggregates incubated overnight as previously.

Blocking Assay:

Article Title: A HER2-Displaying Virus-Like Particle Vaccine Protects from Challenge with Mammary Carcinoma Cells in a Mouse Model
Article Snippet: .. After washing three times with PBS-T, anti-mIgG1-HRP (abcam, ab97240), anti-mIgG2a-HRP (abcam, ab97245), and anti-mIgG2b-HRP (abcam, ab97250) antibodies (1 mg/mL) were diluted 1:3000 in blocking buffer and were added for 1 h at room temperature. .. Plates were washed four times and were developed using OPD substrate (Sigma).

Article Title: Placental extracellular vesicles express active dipeptidyl peptidase IV; levels are increased in gestational diabetes mellitus
Article Snippet: .. Slides were rinsed with distilled water prior to blocking for non-specific antibody binding using 10% Fetal Calf Serum (FCS) (Sigma Aldrich, Gillingham, UK) in PBS-T (PBS with Tween-20) at R/T for 1 h. Sections were incubated overnight at 4°C with 1% FCS and 0.5 µg/ml anti-CD26 primary antibody (Abcam, Cambridge, UK) or non-immune mouse IgG1 (Biolegend, London, UK) in PBS-T. .. Sections were washed in PBS and incubated with anti-mouse IgG secondary antibody (Life Technologies, Paisley, UK) in a humidifying chamber at R/T for 1 h. Slides were washed with 0.01% PBS-T before being stained with DAB (Vector Laboratories, Peterborough, UK).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • pbs t  (Abcam)
    92
    Abcam pbs t
    Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in <t>PBS-T,</t> the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).
    Pbs T, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 470 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs t/product/Abcam
    Average 92 stars, based on 470 article reviews
    Price from $9.99 to $1999.99
    pbs t - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    Image Search Results


    Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).

    Journal: British Journal of Pharmacology

    Article Title: Benzamil sensitive ion channels contribute to volume regulation in canine chondrocytes

    doi: 10.1111/j.1476-5381.2012.02185.x

    Figure Lengend Snippet: Immunohistochemistry and patch-clamp recording reveal the presence of ENaC protein expression and a benzamil and amiloride sensitive cation conductance. Immunofluorescence staining of αENaC (A), βENaC (B) and γENaC (C) in primary cultures of canine articular chondrocytes (first passage cells, 100× objective). Positive immunoreactivity for all subunits was observed although strongest for α- and β-ENaC. The concentration of the primary antibodies was 1 μg·mL −1 (dilution 1:200). The secondary antibody was a goat polyclonal to rabbit IgG (Fc-specific, affinity purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam ab98462). After extensive washes in PBS-T, the nuclei were counterstained with propidium iodide (red fluorescence). Green staining shows the ENaC protein, orange staining is the nuclei and yellow staining is the overlap of the two. (D–F) The negative control was treated identically, but with the omission of primary IgG and shows the red fluorescent nuclear staining with very little background green fluorescence (scale bars for A–F: 10 μm). (G–J) Inside-out patch clamp recordings from canine chondrocytes using inside-out solutions: Tables 1 and 2 . (G) Traces of inside-out low conductance single channel activity at the given membrane potentials. The scale bar horizontal line is 100 ms; vertical line is 500 fA. (H) All-points amplitude histogram for the low conductance channel at −40 mV. (I) Single channel current–voltage curve for the low conductance channel. V rev was −1 ± 5 mV ( n = 5), slope conductance 9 ± 1 pS ( n = 5). (J) Open probability ( P o ) versus time, calculated over successive 0.4 s windows before and during the addition of the ENaC channel inhibitor, amiloride (10 μM). Low conductance single channel P o was reduced by 96 ± 2% ( n = 5).

    Article Snippet: After three washes with PBS-T, the cells were incubated for 2 h with a goat polyclonal secondary antibody to rabbit IgG (Fc-specific, affinity-purified, pre-adsorbed) conjugated to DyLight® 488 (Abcam, Cambridge, MA, USA; ab98462) diluted according to the manufacturer's recommendations (typically diluted 1:200).

    Techniques: Immunohistochemistry, Patch Clamp, Expressing, Immunofluorescence, Staining, Concentration Assay, Affinity Purification, Fluorescence, Negative Control, Activity Assay, Mass Spectrometry