pe conjugated mouse anti integrin β 1 antibody  (R&D Systems)

Journal: Virology Journal

Article Title: Replication kinetics of pathogenic Eurasian orthohantaviruses in human mesangial cells

doi: 10.1186/s12985-024-02517-5

Figure Lengend Snippet: Flow cytometry analysis of surface expression of orthohantaviral entry receptors. CIHGM-1 cells were analyzed for the presence of integrin α v β 3 , integrin β 1 , and CD55 on the cell surface. Plots shown are gated on the viable cell population according to scatter profile and the exclusion of Via-Probe™ Cell Viability Solution. Quadrant statistics (Q1-Q4) indicate the percentage of cells in the respective quadrant

Article Snippet: CIHGM-1 cells were washed with PBS, scraped, and stained with phycoerythrin (PE)-conjugated mouse anti-integrin α V β 3 antibody (clone LM609, Millipore) or PE-conjugated mouse anti-integrin β 1 antibody (clone P5D2, R&D Systems, Minneapolis, MN, USA) together with allophycocyanin (APC)-conjugated anti-CD55 (clone IA10, BD Pharmingen, NJ, USA).

Techniques: Flow Cytometry, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Extracellular matrix remodelling properties of human fibrocytes

doi: 10.1111/j.1582-4934.2011.01344.x

Figure Lengend Snippet: Effects of pre-incubation of fibrocytes with a specific antibody against integrin β 1 or the irrelevant isotype-matched control, and with ligands of the scavenger receptors (poly[I]) and Endo180 (uPA) on cell adhesion to human plasma fibronectin, human collagen I, human collagen IV, human collagen V or human collagen VI. Values are means ± S.D. from five independent experiments. The asterisk indicates statistically significant differences ( P < 0.05) versus medium alone.

Article Snippet: For detection of surface antigens, including the extracellular domains of scavenger and mannose receptors, cells were incubated with one of the following primary antibodies or the isotype-matched control (2–5 μl) for 60 min. in the dark at 4°C: fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD14 (clone HDC14; BioLegend, San Diego, CA, USA); phycoerythrin (PE)-conjugated mouse anti-human CD45 (clone HI30; eBiosciences); peridinin chlorophyll protein-conjugated mouse anti-human CD45 (clone 2D1; BD Biosciences); PE-conjugated mouse anti-human CD34 (clone 8G12; BD Biosciences); PE-conjugated mouse anti-human CD90 (clone Thy-1A1; R&D Systems, Minneapolis, MN, USA); PE-conjugated mouse antibody against human integrin β 1 , also known as CD29, (clone P5D2; R&D Systems); unconjugated rabbit antibody against human Endo180, also known as urokinase-type plasminogen activator (uPA) receptor-associated protein, CD280 or mannose receptor C type 2 (ab70132; Abcam, Cambridge, UK); FITC-conjugated mouse anti-human uPA receptor (American Diagnostica, Stamford, CT, USA); PE-conjugated mouse anti-human CD163 (Clone RM3/1; BioLegend); PE-conjugated mouse antibody against the extracellular domain of CD204, also known as scavenger receptor AI, MSR1 or MSR-AI (clone 351615; R&D Systems); PE-conjugated mouse antibody against human discoidin domain receptor (DDR) 1 (Clone 48B3; Santa Cruz Biotechnology, Santa Cruz, CA, USA); unconjugated mouse antibody against the extracellular domain of DDR2 (clone 290804; IgG 2B , R&D Systems) and unconjugated mouse antibody against CD206, also known as mannose receptor C type 1 or MRC1 (Clone 15–2, IgG 1 ; BioLegend).

Techniques: Incubation, Control, Clinical Proteomics

Journal:

Article Title: CD98 activation increases surface expression and clustering of ? 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

doi: 10.3858/emm.2008.40.3.261

Figure Lengend Snippet: CD98 engagement increases the surface expression of β1 integrin. (A) Freshly cultured MCF-7 cells were dispersed and then incubated with anti-CD98 mAb UM7F8 and secondary Ab (blue line) for 1 h. As negative controls, cells were treated with mouse IgG and secondary Ab (red line). FITC-conjugated anti-CD29 mAb was used to measure the expression level of β1 integrin on the cells treated with mAbs as described above. An FITC-conjugated mouse anti-human lgG was used as the negative control (green line). (B) Cell extracts from MCF-7 cells treated as in (A) were analyzed by Western blotting using anti-β1 integrin and anti-actin mAb.

Article Snippet: R-PE-conjugated mouse anti-β 1 integrin mAb (TDM29) was from Cymbus Biotechnology (Eastleigh, Hampshire, UK).

Techniques: Expressing, Cell Culture, Incubation, Negative Control, Western Blot

Journal:

Article Title: CD98 activation increases surface expression and clustering of ? 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

doi: 10.3858/emm.2008.40.3.261

Figure Lengend Snippet: CD98 cross-linking enhances β1 integrin clustering. (A) MCF-7 cells were incubated with anti-CD98 mAb UM7F8 with or without secondary antibody. To determine whether cross-linking of β1 integrins induces their clustering, cells were treated with anti-β1 integrin mAb 3S3 in the presence of secondary antibody as well. Next, cells treated as described in "Materials and Methods" were analyzed by confocal microscopy. Actin cytoskeleton organization is visualized by staining with phalloidin-FITC whereas β1 integrin clustering with R-PE-conjugated anti-β1 integrin mAb. Images are from a single experiment representative of more than three so performed. Scale bar, 50 µm. Original magnification, × 400. (B) Relative intensities of β1 integrin clusters in Figure 2A were measured with LSM5120 Meta NLO software. Results are values relative to the β1 integrin signal intensity level of untreated controls, designated as 1.

Article Snippet: R-PE-conjugated mouse anti-β 1 integrin mAb (TDM29) was from Cymbus Biotechnology (Eastleigh, Hampshire, UK).

Techniques: Incubation, Confocal Microscopy, Staining, Software

Journal:

Article Title: CD98 activation increases surface expression and clustering of ? 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

doi: 10.3858/emm.2008.40.3.261

Figure Lengend Snippet: Inhibition of FAK/Src kinases with PP2 blocks CD98-induced cell adhesion, but not surface expression and clustering of β1 integrins in MCF-7 cells. (A) The effect of PP2 and/or Mn2+ on FAK phosphorylation in MCF-7 cells treated with anti-CD98 mAb was determined by immunoprecipitaion assay. MCF-7 cells were incubated with anti-CD98 mAb and PP2 (0.2 µM) or a DMSO vehicle control in the presence or absence of 0.5 µM Mn2+ for 1 h and anti-FAK rabbit polyclonal antibody (C-20) was used to immunoprecipitate FAK from extracts of MCF-7 cells. Immunoprecipitates were blotted and probed with anti-phosphotyrosine mAb (clone PY99) and anti-FAK mAb. (B) The effect of PP2 treatment on cell adhesion rate was determined as described in Materials and Methods. In addition, whether addition of 0.5 µM Mn2+ interferes with the effect of PP2 on cell adhesion was determined by the same way. Results are expressed as mean ± SE of values relative to the adhesion rate of mouse IgG-treated controls, designated as 100%. Asterisks show a significant difference from control as follows: *P < 0.05. Additional statistical comparisons are indicated by lines. (C) MCF-7 cells were incubated with anti-CD98 mAb with or without PP2 (0.2 µM) and then analyzed by flow cytometry using FITC-conjugated anti-human β1 integrin mAb. Data represent the mean ± SE of values relative to mean values of fluorescence intensity of mouse IgG-treated controls, designated as 100%. Asterisks show a significant difference from control as follows: *P < 0.05, ***P < 0.001 (D) Confocal microscopy was performed as described in Figure 2 legend to investigate the effect of PP2 (0.2 µM) on CD98-induced clustering of β1 integrins. Scale bar, 50 µm. Original magnification, × 400.

Article Snippet: R-PE-conjugated mouse anti-β 1 integrin mAb (TDM29) was from Cymbus Biotechnology (Eastleigh, Hampshire, UK).

Techniques: Inhibition, Expressing, Phospho-proteomics, Incubation, Control, Flow Cytometry, Fluorescence, Confocal Microscopy

Journal:

Article Title: CD98 activation increases surface expression and clustering of ? 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

doi: 10.3858/emm.2008.40.3.261

Figure Lengend Snippet: The effects of dominant-negative variants of FAK on adhesiveness of MCF-7 cells, surface expression and clustering of β1 integrins. (A) MCF-7 cells were stably transfected with dominant-negative mutant FAK constructs (FRNK, Y397F-FAK) or control vector, pcDNA3. The expression levels of endogenous FAK, FRNK and Y397F-FAK were determined by Western blot analysis using anti-FAK polyclonal antibody. (B) The effect of dominant-negative variants of FAK on CD98-induced cell adhesion rate was determined as described in Materials and Methods. In addition, whether addition of 0.5 µM Mn2+ interferes with the effect of PP2 on cell adhesion was determined by the same way. Results are expressed as in Figure 3 (B). *P < 0.05, **P < 0.01 (C) FAK variants- or mock-transfected MCF-7 cells were treatred with anti-CD98 mAb and secondary antibody and then analyzed for expression of β1 integrin through flow cytometry using FITC-conjugated anti-human β1 integrin. Results are expressed as in Figure 3 (C). Statistical comparisons are indicated by lines. **P < 0.01 (D) Confocal microscopy was performed as described above to investigate the effect of dominant-negative variants of FAK on clustering of β1 integrins. Scale bar, 50 µm. Original magnification, × 400.

Article Snippet: R-PE-conjugated mouse anti-β 1 integrin mAb (TDM29) was from Cymbus Biotechnology (Eastleigh, Hampshire, UK).

Techniques: Dominant Negative Mutation, Expressing, Stable Transfection, Transfection, Construct, Control, Plasmid Preparation, Western Blot, Flow Cytometry, Confocal Microscopy

Journal:

Article Title: CD98 activation increases surface expression and clustering of ? 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

doi: 10.3858/emm.2008.40.3.261

Figure Lengend Snippet: Cytochalasin D or phalloidin treatment inhibits CD98-induced adhesion of MCF-7 cells to fibronectin, but not surface expression and clustering of β1 integrins in MCF-7 cells. (A) MCF-7 cells were incubated with anti-CD98 mAb in the presence or absence of cytochalasin D (4 µM) or phalloidin (10 µM) for 1 h. The effects of DMSO vehicle control are also shown. The effect of cytochalasin D treatment on cell adhesion was determined as described in Materials and Methods. In addition, whether addition of 0.5 µM Mn2+ interferes with the effect of cytochalasin D or phalloidin on cell adhesion was determined by the same way. Results are expressed as Figure 3B. *P < 0.05, **P < 0.01, ***P < 0.001 (B) MCF-7 cells treated with anti-CD98 mAb in the presence or absence of cytochalasin D or phalloidin were analyzed for expression of β1 integrin using flow cytometry. Results are expressed as in Figure 3 (C). **P < 0.01 (C) Confocal microscopy was performed as described above to investigate the effect cytochalasin D or phalloidin of on clustering of β1 integrins. Scale bar, 50 µm. Original magnification, × 400.

Article Snippet: R-PE-conjugated mouse anti-β 1 integrin mAb (TDM29) was from Cymbus Biotechnology (Eastleigh, Hampshire, UK).

Techniques: Expressing, Incubation, Control, Flow Cytometry, Confocal Microscopy