Polyclonal Antibodies

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Western blot analysis of Plexin D1 expression in human endothelial cells (HUVEC) (lanes 1-6). The blots were probed with rabbit polyclonals anti-Plexin D1 (Sema domain) (lanes 1 & 2), anti-Plexin D1 (Cytoplasmic domain) (lanes 3 & 4), and anti-Plexin D1 (a.a. 1635-1647) (lanes 5 & 6). Each antibody was used in the presence of their respective blocking peptide (lanes 2, 4 & 6).
Western blots showing mouse brain (lanes 1 & 4), and Cos-7 cells untransfected (lanes 2 & 5) or transfected with mouse myc-tagged Plexin-A1 (lanes 3 & 6). The blots were probed with either affinity purified anti-Plexin-A1 (PP1471); lanes 1-3) or with mouse monoclonal anti-Myc (lanes 4-6).Immunocytochemical double labeling using anti-Myc mouse monoclonal and anti-Plexin-A1 rabbit polyclonal (PP1471) antibodies in Cos-7 cells mock transfected (A,D) or transfected with Myc-tagged mouse Plexin-A1 construct (B,E). The specificity of the binding in E was demonstrated by using Plexin-A1 peptide (PX1475) in the presence of this anti-Plexin-A1 antibody (C,F).
Western blots showing Cos-7 cells transfected with mouse Myc-tagged Plexin-A1 (lanes 1 & 3), neonatal rat brain (lane 2), or Plexin-A1 immunoprecipitated from Myc-tagged Plexin-A1 transfected cells using anti-Myc (lane 4) or anti-Plexin-A1 (PP1301; lane 5). These blots were probed with either the affinity purified anti-Plexin-A1 (PP1301; lanes 1 & 2) or with mouse monoclonal anti-Myc (lanes 3-5).Immunocytochemical double labeling using anti-Myc mouse monoclonal and anti-Plexin-A1 rabbit polyclonal (PP1301) antibodies in Cos-7 cells mock transfected (A,D) or transfected with Myc-tagged mouse Plexin-A1 construct (B,E). The specificity of the binding in E was demonstrated by using Plexin-A1 peptide (PX1305) in the presence of this anti-Plexin-A1 antibody (C,F).
Bar graph showing rabbit polyclonal anti-Phosphotyrosine (PP2221) binding to a variety of phosphotyrosine containing peptides, but no binding to unphosphorylated peptide (beta-Catenin (a.a. 29-45).Immunocytochemical labeling of phosphotyrosine in control and pervanadate-treated A431 cells. The cells were labeled with rabbit polyclonal anti-Phosphotyrosine (PP2221) and mouse monoclonal anti-Phosphotyrosine (PM3751), then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.
Western blot of human A431 cells treated with Calyculin A (100 nM) for 30 min. The blot was untreated (lane 1) or treated with lambda phosphatase (lanes 2), then probed with anti-Phosphothreonine (PP4641) at 1:1000.Immunocytochemical labeling of phosphothreonine upregulation in control (left) or calyculin A-treated HeLa cells (right). The cells were labeled with rabbit polyclonal anti-Phosphothreonine (PP4641). The antibody was detected using goat anti-rabbit DyLight® 594.
Western blot image of A431 cells untreated (lanes 1 and 3) or treated with pervanadate (lanes 2, 4, 5 & 6). Blots were probed with anti-PDK1 (PP1411) or anti-PDK1 (Tyr-9) (PP1431). The latter was used in the presence of no peptide (lane 4), phospho-PDK1 (Tyr-9) peptide (lane 5), or an unrelated phosphotyrosine peptide (lane 6).Immunocytochemical labeling of PDK1 phosphorylation in control and pervanadate-treated A431 cells. The cells were labeled with mouse monoclonal PDK1 (C-terminal Region) (PM1461), rabbit polyclonal PDK1 (Tyr-9) (PP1431), and PDK1 (N-terminus) (PP1411) antibodies. These antibodies were detected using appropriate secondary antibody conjugated to DyLight® 594.
Western blot analysis of A431 cells. The blot was probed with rabbit polyclonal anti-Paxillin (PP1161) at 1:500 & 1:2000 (lanes 1 & 2) or with mouse monoclonal anti-Paxillin (PM1071) at 1:2000 & 1:5000 (lanes 3 & 4).
Western blots of human PAK6 recombinant protein phosphorylated by ERK2. The blot was exposed to lambda phosphatase (lanes 2 & 6) then probed with anti-PAK6 (N-terminal) (lanes 1-4) or anti-PAK6 (Ser-165) phospho-specific (lanes 5-8). The antibodies were used in the presence of unrelated (lane 3) and PAK6 (N-terminal) (lane 4) peptide or PAK6 (Ser-165) (lane 7) and unrelated phospho-serine (lane 8) peptides, respectively.
Western blot analysis of nNOS expression in adult mouse brain (lanes 1 & 3) and rat GC cells (lanes 2 & 4). The blots were probed with mouse monoclonal anti-nNOS (C-terminal region) at 1:1000 (lanes 1 & 2) or rabbit polyclonal anti-nNOS at 1:250 (lanes 3 & 4).Formalin fixed, citric acid treated parafin sections of adult Rat striatum. Sections were probed with anti-nNOS (NP2141) then anti-Rabbit:HRP before detection using DAB. (Images provided by Carl Hobbs and Dr. Pat Doherty at Wolfson Centre for Age-Related Diseases, King's College London).

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