α human β3 integrin  (Cell Signaling Technology Inc)

Journal: PLoS ONE

Article Title: Adenovirus Gene Transfer to Amelogenesis Imperfecta Ameloblast-Like Cells

doi: 10.1371/journal.pone.0024281

Figure Lengend Snippet: Expression of α v β3 and α v β5 integrins ( A and B ) and heparin sulfate proteoglycans ( C–E ) in AI-WAm cell population and control cells was analyzed by flow cytometry ( A , C , E ) and IHC staining ( B and D ). Cells were incubated with primary anti-α v β3 or anti-α v β5 monoclonal antibodies for detection of corresponding integrin molecules or 10E4 antibody for detection of HSPG side chains (GAG) or anti-human syndecan 4 monoclonal antibody, followed by Alexa 488-conjugated secondary antibody. A and C , top charts: AI-WAm cells; middle charts: RD cells; bottom charts: A549 cells. For AI-WAm cells: P (αvβ3/αvβ5) = 0.75 ; P (Synd4/HSPG) = 0.29 ; For RD cells: P (Synd4/HSPG) = 0.67 ; for α v β3: P (RD/A549) = 0.38 ; for α v β5: P (AI-WAm/A549) = 0.23 ; for syndecan 4: P ( RD /A549) = 0.2 ; for all other differences P <0.05; E. HSPG Ab (10E4) specificity control sample: A549 cells were treated with heparitinase (10 U/ml) for 1 hr at 37°C to remove GAG side chains. Green arrow shows shift of the fluorescence intensity peak resulting from reduction in cell labeling with 10E4 antibody (MFI decrease). Other details are as in Fig. 2D . B and D , scale bars correspond to: 100 µm in top image panels (integrins/AI-WAm, 10× objective), 10 µm (insert, 60× objective) and 50 µm (40× objective) in all other panels. Insert shows syndecan 4 staining image (60×) of A549 cells, clearly demonstrating a polarized intracellular localization of the protein.

Article Snippet: After washes in PBS, the samples were blocked with 10% BSA (Sigma, St. Louis, MO) and incubated overnight at 4°C with one of the following primary antibodies: rabbit polyclonal anti-Amel (Sigma, St. Louis, MO), rabbit polyclonal anti-cytokeratin 14 (ab53115; Abcam, Cambridge, MA); goat polyclonal anti-Enam (C-18; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-human syndecan 4 (Abcam, Cambridge, MA) at 1∶50 dilutions in 3% BSA/PBS or mouse monoclonal anti-hCAR antibody (clone RmcB, Millipore, Billerica, MA), mouse monoclonal anti-human HSPG/GAG 10E4 antibody (F58-10E4, Seikagaku Biobusiness Corp), mouse anti-human integrin α v β3 (LM609 clone) or α v β5 (P1F6 clone) monoclonal antibodies (500 µg/ml) (Millipore, Billerica, MA) at 1∶100 dilutions in 3% BSA/PBS at room temperature for 2 hrs.

Techniques: Expressing, Flow Cytometry, Immunohistochemistry, Incubation, Fluorescence, Labeling, Staining

Journal: PLoS ONE

Article Title: Adenovirus Gene Transfer to Amelogenesis Imperfecta Ameloblast-Like Cells

doi: 10.1371/journal.pone.0024281

Figure Lengend Snippet: A. Differential blocking of gene transfer to AI-WAm cells by integrins. Ad5 RGD shows the highest sensitivity to integrin blocking, while transduction with Ad5-pK7/RGD (G/L) is only partially inhibited. Ad5-pK7 (G/L) gene transfer shows no statistically significant inhibition by integrins. B. Blocking of AI-WAm gene transfer by modified vectors with heparin. Heparin shows a profound dose-dependent blocking effect on transduction with pK7-modified Ads, as opposed to RGD-modified vector. Gray bars (with % values on the top) show percentage of the residual gene transfer level (RLU) resulting from blocking relative to that of unblocked controls (100%) shown by black bar for each fiber-modified vector. All bars represent mean values with standard deviations. All differences were statistically significant except where indicated by asterisk and P values ( P >0.05) on the data bars.

Article Snippet: After washes in PBS, the samples were blocked with 10% BSA (Sigma, St. Louis, MO) and incubated overnight at 4°C with one of the following primary antibodies: rabbit polyclonal anti-Amel (Sigma, St. Louis, MO), rabbit polyclonal anti-cytokeratin 14 (ab53115; Abcam, Cambridge, MA); goat polyclonal anti-Enam (C-18; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-human syndecan 4 (Abcam, Cambridge, MA) at 1∶50 dilutions in 3% BSA/PBS or mouse monoclonal anti-hCAR antibody (clone RmcB, Millipore, Billerica, MA), mouse monoclonal anti-human HSPG/GAG 10E4 antibody (F58-10E4, Seikagaku Biobusiness Corp), mouse anti-human integrin α v β3 (LM609 clone) or α v β5 (P1F6 clone) monoclonal antibodies (500 µg/ml) (Millipore, Billerica, MA) at 1∶100 dilutions in 3% BSA/PBS at room temperature for 2 hrs.

Techniques: Blocking Assay, Transduction, Inhibition, Modification, Plasmid Preparation

Journal: eLife

Article Title: Genetic depletion studies inform receptor usage by virulent hantaviruses in human endothelial cells

doi: 10.7554/eLife.69708

Figure Lengend Snippet: ( a ) Upper panels, total flow cytometry plots of HUVEC and TIME cells stained for endothelial cell markers PECAM and von Willebrand factor (vWF). Medium and lower panels, surface flow cytometry plots of HUVEC and TIME cells stained for PCDH1, β3 integrin, DAF, β1 integrin. ( b ) Surface flow cytometry of wild-type (WT) and knockout (KO) TIME cells stained as above. Histograms of WT cells are shown in gray; single- and double-KO cells are shown in color. ( c ) Western blot analysis of WT TIME cells and KO cells ± cDNA. β-Actin was used as a loading control. Figure 1—source data 1. Original blot of WT TIME cells and KO cells ± cDNA.

Article Snippet: Antibody , α–Human β3 Integrin (Rabbit polyclonal) , Cell Signaling , Cat. # 4702 , WB 1:300.

Techniques: Flow Cytometry, Staining, Knock-Out, Western Blot, Control

Journal: eLife

Article Title: Genetic depletion studies inform receptor usage by virulent hantaviruses in human endothelial cells

doi: 10.7554/eLife.69708

Figure Lengend Snippet:

Article Snippet: Antibody , α–Human β3 Integrin (Rabbit polyclonal) , Cell Signaling , Cat. # 4702 , WB 1:300.

Techniques: Virus, Plasmid Preparation, Expressing, Transduction, Retroviral, Recombinant, Sequencing, Staining, Software, Imaging

Journal: EJNMMI Research

Article Title: [ 99m Tc]Tc-Galacto-RGD 2 integrin α v β 3 -targeted imaging as a surrogate for molecular phenotyping in lung cancer: real-world data

doi: 10.1186/s13550-021-00801-x

Figure Lengend Snippet: Immunohistochemical results among different groups of lung cancer

Article Snippet: Integrin α v β 3 and CXCR4 expression, microvessel density (CD31), and tumour cell proliferation (Ki-67) were detected by incubating the slides with monoclonal antibodies against human integrin α v β 3 (1:200, sc-7312; Santa Cruz Biotechnology, Santa Cruz, California, US), CXCR4 (1:100, ab227767; Abcam, Massachusetts, US), Ki-67 (1:100, ab270650; Abcam), or CD31 (1:50, ab28364; Abcam), respectively, overnight, followed by horseradish peroxidase-conjugated anti-mouse IgG (1:1000, Earth Ox, Millbrae, California, US) with 3′3-diaminobenzidine as the chromogen.

Techniques: Immunohistochemical staining

Journal: EJNMMI Research

Article Title: [ 99m Tc]Tc-Galacto-RGD 2 integrin α v β 3 -targeted imaging as a surrogate for molecular phenotyping in lung cancer: real-world data

doi: 10.1186/s13550-021-00801-x

Figure Lengend Snippet: Immunohistochemical results of lung cancer and benign disease

Article Snippet: Integrin α v β 3 and CXCR4 expression, microvessel density (CD31), and tumour cell proliferation (Ki-67) were detected by incubating the slides with monoclonal antibodies against human integrin α v β 3 (1:200, sc-7312; Santa Cruz Biotechnology, Santa Cruz, California, US), CXCR4 (1:100, ab227767; Abcam, Massachusetts, US), Ki-67 (1:100, ab270650; Abcam), or CD31 (1:50, ab28364; Abcam), respectively, overnight, followed by horseradish peroxidase-conjugated anti-mouse IgG (1:1000, Earth Ox, Millbrae, California, US) with 3′3-diaminobenzidine as the chromogen.

Techniques: Immunohistochemical staining

Journal: EJNMMI Research

Article Title: [ 99m Tc]Tc-Galacto-RGD 2 integrin α v β 3 -targeted imaging as a surrogate for molecular phenotyping in lung cancer: real-world data

doi: 10.1186/s13550-021-00801-x

Figure Lengend Snippet: RGD-avid lesion in a patient with lung adenocarcinoma, consistent with α v β 3 expression which confirmed by IHC, flow cytometry and western blot. a – c [ 99m Tc]Tc-Galacto-RGD 2 image. d , e IHC showed higher expression of α v β 3 in the tumour cells and higher density of micro-vessel (CD31). f Flow cytometry showed higher expression of integrin β 3 in tumour tissue compared with normal lung tissue. g Western blot showed higher integrin β 3 expression in primary tumour tissue compared with normal lung tissue

Article Snippet: Integrin α v β 3 and CXCR4 expression, microvessel density (CD31), and tumour cell proliferation (Ki-67) were detected by incubating the slides with monoclonal antibodies against human integrin α v β 3 (1:200, sc-7312; Santa Cruz Biotechnology, Santa Cruz, California, US), CXCR4 (1:100, ab227767; Abcam, Massachusetts, US), Ki-67 (1:100, ab270650; Abcam), or CD31 (1:50, ab28364; Abcam), respectively, overnight, followed by horseradish peroxidase-conjugated anti-mouse IgG (1:1000, Earth Ox, Millbrae, California, US) with 3′3-diaminobenzidine as the chromogen.

Techniques: Expressing, Flow Cytometry, Western Blot