rabbit polyclonal anti β4 integrin antibody  (Bioss)

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Mutual localization of the α6 and β4 integrin subunit and a presence of α6β4 heterodimer revealed by SIM and PLA. ( a ) α6 (red) and β4 (green) are localized in the plasma membrane overlying the acrosomal cap (for details see panel c , white arrows), apical hook and equatorial segment in an intact sperm head. ( b ) PLA confirmed the presence of the α6β4 heterodimer in depicted locations ( a ). ( c ) SIM dual-colour imaging showing merged image of α6 (red) and β4 (green) localized in the plasma membrane (white arrows). ( d ) Huygens software was used for a better visualization of the colocalization area (yellow). Colocalization maps are based on Pearson’s correlation coefficient. Nucleus is visualized with Dapi (blue). Scale bar represents 1 µm ( a – c ) and 2 µm ( d ).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Imaging, Software

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Gene expression of β4 integrin in the cell-type fractions. Cq value of the gene is normalized by reference gene Rps2 . Numbers > 1 are considered as strongly expressed in the individual cell-types.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: ( a ) Primers designing and ( b ) agarose gel electrophoresis of PCR products involving in the cytoplasmic domain of β4 integrin. 1–5 PCR products amplified by primer pairs (pp) in mRNA sperm samples after elutriation.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Agarose Gel Electrophoresis, Amplification

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Western blot immunodetection of the β4 integrin in protein extract from mouse epididymal sperm with mouse monoclonal anti-β4 integrin (sc-13543) antibody; (1) antibody reaction in protein extract from mouse sperm, (2) negative control with mouse IgG; detection of 200 kDa protein band corresponds to β4 integrin (black arrow), possible high molecular weight isoforms (white arrows), non-specific reaction (grey arrows).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Western Blot, Immunodetection, Negative Control, Molecular Weight

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Mutual localization of α3 and β1 integrin subunit and a presence of α3β1 heterodimer revealed by SIM and PLA. ( a ) α3 (green) and β1 (red) are localized in the acrosomal cap area of intact sperm head, ( b ) PLA confirmed a presence of the α3β1 heterodimer. ( c ) SIM depicted their mutual localization in same structures. ( d ) Huygens software was used for better visualization of colocalization area (yellow) of α3 and β1. Colocalization maps are based on Pearson’s correlation coefficient. Nucleus is visualized with Dapi (blue). Scale bar represents 1 μm ( a – c ) and 2 μm ( d ).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Software

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Docking of α/β integrin N-terminal domains. ( a , b ) An example of resting state structure of extracellular domains of the α3β1 integrin as obtained from I-TASSER homology modelling. As expected from the crystal structures available for threading by I-TASSER the overall architecture of the resulting model is similar to αXβ2 (PDB ID 4neh; ) or αIIbβ3 (PDB ID 3fcs; ) heterodimer structures. The interacting region of N-terminal domains is highlighted by the black rectangle. Panel ( a ) shows the α3 subunit on top of the β1 domains, while panel ( b ) represents the same structure rotated by 90 degrees orienting both the N-terminal domains to the front of the image. ( c – f ) The predicted arrangement of heterodimers formed by N-terminal domains of integrins α3β1 ( c ), α6β1 ( d ), α3β4 ( e ), and α6β4 ( f ) from the flexible side chain docking of the domains by ClusPro. In the case of α3β4 ( f ) two binding modes are shown. The predicted most stable dimer with the β4 domain (orange) interacting through the NV residues homologous with the typical Arg/Lys finger of β domains is compared to an alternative, less stable binding mode, employing the RPEK sequence (grey). Both binding modes are similar and suggest that the α3β4 complex adopts different domain orientation compared to the remaining α3β1, α6β1, and α6β4 cases. The α3 domains are shown in green, β1 in blue, α6 in red, and β4 in orange.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Binding Assay, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Graphical summary of integrin heterodimers and their localization in intact sperm head. ( a ) Localization of integrin heterodimers as follows: α3β1, α6β1, α6β4–plasma membrane (PM) overlaying the apical acrosome (AA), α6β1, α6β4–sperm hook (H); α3β–outer acrosomal membrane of apical acrosome (AA); α6β4–inner acrosomal membrane and equatorial segment (ES) up to posterior post-acrosomal region (PAR). ( b ) Schematic depiction of integrin subunits anchored in the membrane, dynamics between α3 and α6 with β1 and β subunits interaction with cytoskeleton.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Model of activated states of α/β integrin heterodimers. A probable arrangement of integrin heterodimers in the open state was derived from the N-terminal domain complexes predicted by docking (see ) by a structural superposition of the existing crystal structure of αIIBβ3 in the open state (PDB ID 3fcu; ). The modelling suggests that while for the α3β1, α6β1 ( a ), and α6β4 ( b ) the integrins are involved in a cis interaction adopting the expected conformation with the membrane proximal domains separated [ , , , ], the orientation of the α3β4 N-terminal domains would lead to a complex with α and β subunits pointing in nearly opposite directions ( b ) supporting the possible trans interaction (see for more details).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Diagram of sperm specific integrin subunits and heterodimers. The solid lines show proven integrin heterodimers α3β1, α6β1 and α6β4; the arrows show predicted α3 and α6 dynamics with β1; α3 and β4 cis interaction are energetically unstable in contract to trans interaction. Heterodimers between αV and β1/β3 are not known.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques:

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: β4 integrin is highly expressed in human osteosarcoma cell lines and tumor samples. A, β4 and β1 integrin expression were determined by Western blot analysis and β4 integrin was also analyzed by Northern blot analysis in 4 osteosarcoma (U2OS, SaOS, G292, and HOS), 2 rhabdomyosarcoma (RD and Rh18), and 2 Ewing’s sarcoma (RDES and TC32) cell lines. B, expression of β4, β1, β3, and α6 integrins was determined by Western blot analysis in the non-metastatic human osteosarcoma cell line HOS and the highly metastatic cell line MNNG-HOS. C, tissue array analysis of β4 integrin expression was performed on human osteosarcoma samples by immunohistochemistry.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Expressing, Western Blot, Northern Blot, Immunohistochemistry

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: Knockdown of β4 integrin expression results in a clumpy cell morphology in 3D co-cultures. A, MNNG-HOS (Luc) cells were infected with lentivirus expressing control-shRNA or β4 integrin-shRNA. Expression β4 and α6 integrins was analyzed by Western blot analysis in control-shRNA or β4 integrin-shRNA infected cells. B, the cell surface expression of β4 integrin was evaluated by FACS in control and β4 integrin knockdown cells. Normal IgG was used as a negative control (figure not shown). C, the surface expression and localization of β4 integrin were examined by immunofluorescent staining using same antibody as above in control and knockdown cells. D, MNNG-HOS (Luc) cells infected with lentivirus of control-shRNA (left panel) or β4 integrin-shRNA (right panel) are shown on top as traditional 2D images. Below, are ostoesarcoma cells that were labeled with CellTracker Green CMTPX dye (green). NIH3T3 fibroblasts were labeled separately with CellTracker Orange CMRA dye (red). All cells were then labeled with Hoechst 33342 nuclear dye (blue) and co-cultured in a nanofiber-based matrix labeled with Cy5 dye (white) containing Cultrex basement membrane extract. The lower right panels show merged views.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Knockdown, Expressing, Infection, Control, shRNA, Western Blot, Negative Control, Staining, Labeling, Cell Culture, Membrane

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: Control-shRNA and β4 integrin-shRNA infected MNNG-HOS (Luc) cells were injected into the tail vein of RAG knockout mice. These mice then underwent luminescent imaging. Suppression of β4 integrin expression by shRNA led to significant reduction of luminescent intensity in the lungs of mice 50 days after injection of cells, shown graphically (A) with representative lung images at time of sacrifice (B). C, quantitation of luminescence for animals in (A). D, Kaplan-Meier survival curves for MNNG-HOS control-shRNA or β4-shRNA cells.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Control, shRNA, Infection, Injection, Knock-Out, Imaging, Expressing, Quantitation Assay

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: Effect of dominant negative β4 integrin expression on primary tumor growth and experimental metastases in vivo . A, disruption of β4 integrin function by transfection of mutant dominant negative β4 integrin. MNNG-HOS cells were transfected with empty vector (EV) and dominant-negative β4 integrin (Y1494F). After batch selection with medium containing G418, cells were lysed and subjected to Western blot analysis of β4 integrin expression (top panel), and immunoprecipititated with β4 integrin antibody overnight followed by Western blot for phospho-tyrosine and β4 integrin (lower panel). B, the cell surface expression of mutant β4 integrin was measured by FACS in mutant β4 integrin (Y1494F) transfected cells. C, empty vector (EV)- and dominant negative β4 integrin (β4-M)-transfected MNNG-HOS (luc) cells were injected into the gastrocnemius muscle of SCID/BEIGE mice and assayed for growth of primary tumors by luminescent imaging (left panel) and quantitation of luminescence signal (right panel). D, empty vector (EV)- and dominant negative β4 integrin (β4M)-transfected MNNG-HOS (luc) cells were injected into the tail vein of RAG knockout mice. Luminescence was quantified weekly with mean intensity for each group plotted over time. E, Kaplan-Meier survival curves showing percent survival for each cell line over time.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Dominant Negative Mutation, Expressing, In Vivo, Disruption, Transfection, Mutagenesis, Plasmid Preparation, Selection, Western Blot, Injection, Imaging, Quantitation Assay, Knock-Out

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: β4 integrin interacts with ezrin. A and B, cell lysates were immunoprecipitated (IP) with an anti-ezrin antibody overnight followed by Western blot analysis for β4 and ezrin using the indicated antibodies (WB). C, purified recombinant proteins for BSA, ezrin-N-terminal region (residues 1–297)(Ez-NT), GST, and GST-ezrin-C-terminal region (residues 280–585)(GST-Ez-CT) were immobilized onto glutathione-sepharose beads and incubated overnight with lysates (input) from SaOS cells or in vitro synthesized β4 integrin. The bound fractions were washed and subjected to SDS-PAGE, followed by immunoblotting with anti-β4 integrin antibody.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Immunoprecipitation, Western Blot, Purification, Recombinant, Incubation, In Vitro, Synthesized, SDS Page

Journal: Oncogene

Article Title: Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

doi: 10.1038/onc.2009.206

Figure Lengend Snippet: Both ezrin expression and function are required for maintenance of β4 integrin protein expression. A, The expression of ezrin, β1, β3, and β4 integrin, were analyzed by Western blot in K12, K7M2, and K7M2-ezrin antisense cell lines. B, K7M2 and HOS transfected with control (C) or ezrin (Ez) or β4 siRNA. After 3 days, cells were lysed and subjected to Western blot analysis for ezrin and β4 integrin expression. C, β4 integrin expression was analyzed by Western blot in empty vector-GFP and mutant ezrin (T567A) line clones derived from the parental K7M2 cell line. D, K7M2-ezrin antisense cell line clones 13, 1.46 and 1.52 were treated with the proteosome inhibitor, MG132 (50 mM), for 6 h and then analyzed by Western blot for ezrin and β4 integrin expression. E, quantitative RT-PCR analysis for steady-state levels of β4 integrin mRNA in parental K7M2 cells relative to the K7M2-ezrin antisense 1.46 cell line. Samples were normalized to GAPDH mRNA. Mean normalized fold expression is shown from duplicate, independently isolated RNA samples for each cell line.

Article Snippet: Rabbit polyclonal β4 integrin primary antibody (sc9090, Santa Cruz Biotechnology) at 1:50 dilution was added overnight followed by biotinylated polyclonal goat anti-rabbit immunoglobulin secondary antibody (Dako) at 1:100 dilution for 1 hour.

Techniques: Expressing, Western Blot, Transfection, Control, Plasmid Preparation, Mutagenesis, Clone Assay, Derivative Assay, Quantitative RT-PCR, Isolation