goat anti rabbit igg h l highly cross adsorbed secondary antibody  (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
  • 97
    Name:
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody
    Description:
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody for IF ICC Flow
    Catalog Number:
    A11034
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Antibodies and Secondary Detection|Cell Analysis|Secondary Detection
    Buy from Supplier


    Structured Review

    Thermo Fisher goat anti rabbit igg h l highly cross adsorbed secondary antibody
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody for IF ICC Flow
    https://www.bioz.com/result/goat anti rabbit igg h l highly cross adsorbed secondary antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg h l highly cross adsorbed secondary antibody - by Bioz Stars, 2021-04
    97/100 stars

    Images

    Related Articles

    other:

    Article Title: Common cytotoxic chemotherapeutics induce epithelial-mesenchymal transition (EMT) downstream of ER stress
    Article Snippet: Alexa Fluor 488 goat anti-rabbit IgG #A11034 and Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Incubation:

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Article Title: Formation and spreading of TDP-43 aggregates in cultured neuronal and glial cells demonstrated by time-lapse imaging
    Article Snippet: Immunocytochemistry Cells were fixed with 4% paraformaldehyde in phosphate buffered saline (PBS), permeabilized with 100% methanol, washed with PBS, and immunostained overnight at 4°C with the following primary antibodies at 1:200 dilutions; mouse monoclonal TuJ1 (R & D systems, Minneapolis, MN, USA), rabbit anti-phosphoTDP-43 (pSer409/410) (Cosmo Bio), rabbit anti-TDP-43 C-terminus (Abcam, Cambridge, MA, USA), rabbit anti-GFP (Abcam), rabbit anti-GFAP (DAKO, Glostrup, Denmark), rabbit anti-ubiquitin (DAKO) and rabbit anti-CNP (Cell Signaling Technology, Danvers, MA, USA). .. The cells were then incubated with Alexa Fluor 488- or 633-conjugated goat anti-rabbit or anti-mouse antibodies (Thermo Fisher) at 1:400 dilutions for 45 min at room temperature, followed by incubation for 15 min with 2 μg/mL Hoechst 33342 (Thermo Fisher). ..

    Labeling:

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Staining:

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Transduction:

    Article Title: Role of lipid rafts in E-cadherin- and HGF-R/Met-mediated entry of Listeria monocytogenes into host cells
    Article Snippet: .. Reagents and antibodies Alexa Fluor 488 cholera toxin subunit B, Alexa Fluor 488–, 546–, and 647–conjugated phalloidin, Alexa Fluor 488– and 546–conjugated goat anti–rabbit and goat anti–mouse antibodies were purchased from Molecular Probes. mAbs anti–E-cadherin (clone HECD-1) was purchased from R & D systems, anti–α-catenin (clone 5) was purchased from Transduction Laboratories. ..

    Immunofluorescence:

    Article Title: Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes
    Article Snippet: .. Indirect immunofluorescence was performed using secondary antibodies conjugated with Alexa-647 anti-Mouse (Invitrogen, Cat. A-21236), Alexa-555 anti-Rat (Invitrogen, Cat. A-21434) and Alexa-488 anti-Rabbit (Invitrogen, Cat. A-11034). .. 10 mg/ml Hoechst 33342 stock solution was purchased from Life Technologies (Cat. H3570).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher alexa 488 anti rabbit
    Efficacy of fluorophore inactivation and preservation of tissue integrity. ( A ) Exemplary image of a human tonsil stained with PCNA-Alexa 488 that underwent 0, 15, 30 or 60 min of fluorophore inactivation. ( B ) Effect of bleaching duration on the distribution of anti-PCNA-Alexa 488 staining intensities for samples used in ( A ). The distribution is computed from mean values for the fluorescence intensities across all cells in the image that were successfully segmented. The gray band denotes the range of background florescence intensities (below 6.2 in log scale). ( C ) Effect of bleaching duration on mean intensity for nine antibodies conjugated to Alexa fluor 488, efluor 570 or Alexa fluor 647. Intensities were determined as in ( B ). The gray band denotes the range of background florescence intensities. ( D ) Impact of t-CyCIF cycle number on tissue integrity for four exemplary tissue cores. Nuclei present in the first cycle are labeled in red and those present after the 10th cycle are in green. The numbers at the bottom of the images represent nuclear counts in cycle 1 (red) and cycle 10 (green), respectively. ( E ) Impact of t-CyCIF cycle number on the integrity of a TMA containing 48 biopsies obtained from 16 different healthy and tumor tissues (see Materials and methods for TMA details) stained with 10 rounds of t-CyCIF. The number of nuclei remaining in each core was computed relative to the starting value; small fluctuations in cell count explain values  >  1.0 and arise from errors in image segmentation. Data for six different breast cores is shown to the right. ( F ) Nuclear staining of a melanoma specimen subjected to 20 cycles of t-CyCIF emphasizes the preservation of tissue integrity (22 ± 4%). ( G ) Selected images of the specimen in ( F ) from cycles 0, 5, 15 and 20.
    Alexa 488 Anti Rabbit, 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/alexa 488 anti rabbit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa 488 anti rabbit - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    Efficacy of fluorophore inactivation and preservation of tissue integrity. ( A ) Exemplary image of a human tonsil stained with PCNA-Alexa 488 that underwent 0, 15, 30 or 60 min of fluorophore inactivation. ( B ) Effect of bleaching duration on the distribution of anti-PCNA-Alexa 488 staining intensities for samples used in ( A ). The distribution is computed from mean values for the fluorescence intensities across all cells in the image that were successfully segmented. The gray band denotes the range of background florescence intensities (below 6.2 in log scale). ( C ) Effect of bleaching duration on mean intensity for nine antibodies conjugated to Alexa fluor 488, efluor 570 or Alexa fluor 647. Intensities were determined as in ( B ). The gray band denotes the range of background florescence intensities. ( D ) Impact of t-CyCIF cycle number on tissue integrity for four exemplary tissue cores. Nuclei present in the first cycle are labeled in red and those present after the 10th cycle are in green. The numbers at the bottom of the images represent nuclear counts in cycle 1 (red) and cycle 10 (green), respectively. ( E ) Impact of t-CyCIF cycle number on the integrity of a TMA containing 48 biopsies obtained from 16 different healthy and tumor tissues (see Materials and methods for TMA details) stained with 10 rounds of t-CyCIF. The number of nuclei remaining in each core was computed relative to the starting value; small fluctuations in cell count explain values  >  1.0 and arise from errors in image segmentation. Data for six different breast cores is shown to the right. ( F ) Nuclear staining of a melanoma specimen subjected to 20 cycles of t-CyCIF emphasizes the preservation of tissue integrity (22 ± 4%). ( G ) Selected images of the specimen in ( F ) from cycles 0, 5, 15 and 20.

    Journal: eLife

    Article Title: Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes

    doi: 10.7554/eLife.31657

    Figure Lengend Snippet: Efficacy of fluorophore inactivation and preservation of tissue integrity. ( A ) Exemplary image of a human tonsil stained with PCNA-Alexa 488 that underwent 0, 15, 30 or 60 min of fluorophore inactivation. ( B ) Effect of bleaching duration on the distribution of anti-PCNA-Alexa 488 staining intensities for samples used in ( A ). The distribution is computed from mean values for the fluorescence intensities across all cells in the image that were successfully segmented. The gray band denotes the range of background florescence intensities (below 6.2 in log scale). ( C ) Effect of bleaching duration on mean intensity for nine antibodies conjugated to Alexa fluor 488, efluor 570 or Alexa fluor 647. Intensities were determined as in ( B ). The gray band denotes the range of background florescence intensities. ( D ) Impact of t-CyCIF cycle number on tissue integrity for four exemplary tissue cores. Nuclei present in the first cycle are labeled in red and those present after the 10th cycle are in green. The numbers at the bottom of the images represent nuclear counts in cycle 1 (red) and cycle 10 (green), respectively. ( E ) Impact of t-CyCIF cycle number on the integrity of a TMA containing 48 biopsies obtained from 16 different healthy and tumor tissues (see Materials and methods for TMA details) stained with 10 rounds of t-CyCIF. The number of nuclei remaining in each core was computed relative to the starting value; small fluctuations in cell count explain values  >  1.0 and arise from errors in image segmentation. Data for six different breast cores is shown to the right. ( F ) Nuclear staining of a melanoma specimen subjected to 20 cycles of t-CyCIF emphasizes the preservation of tissue integrity (22 ± 4%). ( G ) Selected images of the specimen in ( F ) from cycles 0, 5, 15 and 20.

    Article Snippet: Indirect immunofluorescence was performed using secondary antibodies conjugated with Alexa-647 anti-Mouse (Invitrogen, Cat. A-21236), Alexa-555 anti-Rat (Invitrogen, Cat. A-21434) and Alexa-488 anti-Rabbit (Invitrogen, Cat. A-11034).

    Techniques: Preserving, Staining, Fluorescence, Labeling, Cell Counting

    Cells expressing GFP-Kin1p were starved for 24 h, deciliated, and allowed to regenerate cilia. At various times, samples of cells were processed for immunofluorescence using anti-GFP antibodies plus secondary antibodies coupled to Cy3 and TAP952 anti-tubulin antibodies plus secondary antibodies coupled to FITC. Pairs of corresponding GFP (A, C, E, and G) and tubulin (B, D, F, and H) images are shown for individual cells. Bars, 15 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Kinesin-II Is Preferentially Targeted to Assembling Cilia and Is Required for Ciliogenesis and Normal Cytokinesis in Tetrahymena V⃞

    doi:

    Figure Lengend Snippet: Cells expressing GFP-Kin1p were starved for 24 h, deciliated, and allowed to regenerate cilia. At various times, samples of cells were processed for immunofluorescence using anti-GFP antibodies plus secondary antibodies coupled to Cy3 and TAP952 anti-tubulin antibodies plus secondary antibodies coupled to FITC. Pairs of corresponding GFP (A, C, E, and G) and tubulin (B, D, F, and H) images are shown for individual cells. Bars, 15 μm.

    Article Snippet: Secondary antibodies were goat anti-mouse FITC (Sigma), goat anti-rabbit-Cy3, and goat-anti-rabbit-FITC (Zymed, San Francisco, CA) conjugates, and all were used at 1:100 dilution.

    Techniques: Expressing, Immunofluorescence

    Wild-type (A–F, M, and N) and GFP-kin1p rescued (G–L, O, and P) cells were isolated from exponentially growing cultures and processed for confocal analysis using anti-GFP antibodies and TAP952 anti-tubulin antibodies. GFP and tubulin were detected using secondary antibodies coupled to Cy3 and FITC, respectively. For individual cells corresponding images of GFP (A, C, E, G, I, K, M, and O) and tubulin (B, D, F, H, J, L, N, and P) are shown. (A and B) Negative control cell. Some background staining with GFP antibodies is present in the cell body (A). (C–F) higher magnifications of the boxes shown in panels A and B. (G and H) Nondividing cell expressing GFP-Kin1p. Note weak staining of cilia by anti-GFP antibodies. Some cilia shown in boxed areas are labeled more strongly by anti-GFP antibodies (G). These cilia are shorter and label more uniformly by the TAP952 antibodies (H). (I–L) Higher magnifications of the boxes shown in G and H. Arrowheads show shorter immature cilia, which label uniformly with TAP952 antibodies and also show more of GFP signal. Stars indicate mature cilia in which the TAP952 labeling is restricted to ciliary tips, and there is relatively less GFP signal. (M and N) Negative control dividing cell. Two oral apparatuses are present. (O and P) Dividing cell expressing GFP-Kin1p. The oral cilia are labeled heavily by anti-GFP antibodies (O) and uniformly by TAP952 antibodies (P). Note relatively weak staining for GFP in the oral apparatus of a nondividing cell (G). Bars: A and M, 15 μm; C, 1 μm. OA, oral apparatus; LM, longitudinal microtubule bundle.

    Journal: Molecular Biology of the Cell

    Article Title: Kinesin-II Is Preferentially Targeted to Assembling Cilia and Is Required for Ciliogenesis and Normal Cytokinesis in Tetrahymena V⃞

    doi:

    Figure Lengend Snippet: Wild-type (A–F, M, and N) and GFP-kin1p rescued (G–L, O, and P) cells were isolated from exponentially growing cultures and processed for confocal analysis using anti-GFP antibodies and TAP952 anti-tubulin antibodies. GFP and tubulin were detected using secondary antibodies coupled to Cy3 and FITC, respectively. For individual cells corresponding images of GFP (A, C, E, G, I, K, M, and O) and tubulin (B, D, F, H, J, L, N, and P) are shown. (A and B) Negative control cell. Some background staining with GFP antibodies is present in the cell body (A). (C–F) higher magnifications of the boxes shown in panels A and B. (G and H) Nondividing cell expressing GFP-Kin1p. Note weak staining of cilia by anti-GFP antibodies. Some cilia shown in boxed areas are labeled more strongly by anti-GFP antibodies (G). These cilia are shorter and label more uniformly by the TAP952 antibodies (H). (I–L) Higher magnifications of the boxes shown in G and H. Arrowheads show shorter immature cilia, which label uniformly with TAP952 antibodies and also show more of GFP signal. Stars indicate mature cilia in which the TAP952 labeling is restricted to ciliary tips, and there is relatively less GFP signal. (M and N) Negative control dividing cell. Two oral apparatuses are present. (O and P) Dividing cell expressing GFP-Kin1p. The oral cilia are labeled heavily by anti-GFP antibodies (O) and uniformly by TAP952 antibodies (P). Note relatively weak staining for GFP in the oral apparatus of a nondividing cell (G). Bars: A and M, 15 μm; C, 1 μm. OA, oral apparatus; LM, longitudinal microtubule bundle.

    Article Snippet: Secondary antibodies were goat anti-mouse FITC (Sigma), goat anti-rabbit-Cy3, and goat-anti-rabbit-FITC (Zymed, San Francisco, CA) conjugates, and all were used at 1:100 dilution.

    Techniques: Isolation, Negative Control, Staining, Expressing, Labeling

    Chemotherapeutic drugs and thapsigargin treatment activate EMT ( A ) Fluorescence staining for E-cadherin and Vimentin in A549 cells. Cells were plated on chamber slides and either treated with vehicle alone or with known chemotherapeutic drugs with conc. indicated in Figure 4 . After 48 hrs of treatment cells were fixed and stained for EMT markers. i, iii, v, vii, ix and xi: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii, iv, vi, viii, x and xii: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c, e, g, i and k: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d, f, h, j and l: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. ( B ) A549 cells were prepared as described in A and F-actin was detected with Alexa Fluor 568 Phalloidin (red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows. ( C ) Fluorescence staining for F-actin in A549 (i and ii) and H358 (iii and iv) cells. Cells were either treated with vehicle alone or with 0.5 μM of thapsigargin for 30 min and without thapsigargin in medium for 24 hr. After 24 hrs of treatment cells were trypsinized and plated on chamber slides and stained for F-actin (Alexa Fluor 568 Phalloidin; red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows.

    Journal: Oncotarget

    Article Title: Common cytotoxic chemotherapeutics induce epithelial-mesenchymal transition (EMT) downstream of ER stress

    doi: 10.18632/oncotarget.15150

    Figure Lengend Snippet: Chemotherapeutic drugs and thapsigargin treatment activate EMT ( A ) Fluorescence staining for E-cadherin and Vimentin in A549 cells. Cells were plated on chamber slides and either treated with vehicle alone or with known chemotherapeutic drugs with conc. indicated in Figure 4 . After 48 hrs of treatment cells were fixed and stained for EMT markers. i, iii, v, vii, ix and xi: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii, iv, vi, viii, x and xii: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c, e, g, i and k: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d, f, h, j and l: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. ( B ) A549 cells were prepared as described in A and F-actin was detected with Alexa Fluor 568 Phalloidin (red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows. ( C ) Fluorescence staining for F-actin in A549 (i and ii) and H358 (iii and iv) cells. Cells were either treated with vehicle alone or with 0.5 μM of thapsigargin for 30 min and without thapsigargin in medium for 24 hr. After 24 hrs of treatment cells were trypsinized and plated on chamber slides and stained for F-actin (Alexa Fluor 568 Phalloidin; red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows.

    Article Snippet: Alexa Fluor 488 goat anti-rabbit IgG #A11034 and Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Techniques: Fluorescence, Staining

    ER stress induces EMT ( A ) Western blot analysis of EMT markers in A549 and H358 cells. Cells were either treated with vehicle alone or with 0.5μM of thapsigargin for 30min and without thapsigargin in medium for 24hr. Cell lysates were prepared and western blot analysis of EMT markers Vimentin and E-cadherin was performed. ( B ) Fluorescence staining for E-cadherin and Vimentin in A549 and H358 cells. Cells were either treated with vehicle alone or with 0.5 μM of thapsigargin for 30 min and without thapsigargin in medium for 24 hr. After 24 hrs of treatment cells were trypsinized and plated on chamber slides and stained for EMT markers. i, and iii: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii and iv: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a and c: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b and d: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. ( C ) Determination of concentration of thapsigargin inducing ER stress response and EMT. A549 cells were treated with indicated concentration of thapsigargin for 24 hrs. A549 cell lysates were prepared and analyzed for ER stress and EMT markers.

    Journal: Oncotarget

    Article Title: Common cytotoxic chemotherapeutics induce epithelial-mesenchymal transition (EMT) downstream of ER stress

    doi: 10.18632/oncotarget.15150

    Figure Lengend Snippet: ER stress induces EMT ( A ) Western blot analysis of EMT markers in A549 and H358 cells. Cells were either treated with vehicle alone or with 0.5μM of thapsigargin for 30min and without thapsigargin in medium for 24hr. Cell lysates were prepared and western blot analysis of EMT markers Vimentin and E-cadherin was performed. ( B ) Fluorescence staining for E-cadherin and Vimentin in A549 and H358 cells. Cells were either treated with vehicle alone or with 0.5 μM of thapsigargin for 30 min and without thapsigargin in medium for 24 hr. After 24 hrs of treatment cells were trypsinized and plated on chamber slides and stained for EMT markers. i, and iii: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii and iv: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a and c: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b and d: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. ( C ) Determination of concentration of thapsigargin inducing ER stress response and EMT. A549 cells were treated with indicated concentration of thapsigargin for 24 hrs. A549 cell lysates were prepared and analyzed for ER stress and EMT markers.

    Article Snippet: Alexa Fluor 488 goat anti-rabbit IgG #A11034 and Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Techniques: Western Blot, Fluorescence, Staining, Concentration Assay

    Formation of cytoplasmic TDP-43 aggregates is accelerated by proteasome inhibition in neuronally-differentiated 1464R cells. Differentiated 1464R neuronal cells were transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (DsR-(WT+CTF) (red) for 48 hrs and treated with DMSO ( a–c ) or MG-132 ( d–f ) for an additional 24 hrs. Fixed cells were immunostained for TuJ1 ( a–f ), TDP-43 ( a, d ), phosphorylated TDP-43 (pSer409/Ser410) (phosphoTDP-43) ( b, e ) and ubiquitin ( c, f ) with Alexa Fluor 488- or 633-conjugated secondary antibodies, and counterstained with Hoechst 33342 (H33342; blue). In MG-132-treated TuJ1-positive neurons ( a–c ), cytoplasmic aggregates (arrows) are strongly immunoreactive for TDP-43 ( d ), phosphoTDP-43 ( e ) and ubiquitin ( f ). Scale bar = 20 μm.

    Journal: PLoS ONE

    Article Title: Formation and spreading of TDP-43 aggregates in cultured neuronal and glial cells demonstrated by time-lapse imaging

    doi: 10.1371/journal.pone.0179375

    Figure Lengend Snippet: Formation of cytoplasmic TDP-43 aggregates is accelerated by proteasome inhibition in neuronally-differentiated 1464R cells. Differentiated 1464R neuronal cells were transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (DsR-(WT+CTF) (red) for 48 hrs and treated with DMSO ( a–c ) or MG-132 ( d–f ) for an additional 24 hrs. Fixed cells were immunostained for TuJ1 ( a–f ), TDP-43 ( a, d ), phosphorylated TDP-43 (pSer409/Ser410) (phosphoTDP-43) ( b, e ) and ubiquitin ( c, f ) with Alexa Fluor 488- or 633-conjugated secondary antibodies, and counterstained with Hoechst 33342 (H33342; blue). In MG-132-treated TuJ1-positive neurons ( a–c ), cytoplasmic aggregates (arrows) are strongly immunoreactive for TDP-43 ( d ), phosphoTDP-43 ( e ) and ubiquitin ( f ). Scale bar = 20 μm.

    Article Snippet: The cells were then incubated with Alexa Fluor 488- or 633-conjugated goat anti-rabbit or anti-mouse antibodies (Thermo Fisher) at 1:400 dilutions for 45 min at room temperature, followed by incubation for 15 min with 2 μg/mL Hoechst 33342 (Thermo Fisher).

    Techniques: Inhibition, Transduction