polyclonal anti alk2 antibody  (R&D Systems)

Journal: Journal of Neuroscience

Article Title: Bone Morphogenetic Protein Signaling in the Developing Telencephalon Controls Formation of the Hippocampal Dentate Gyrus and Modifies Fear-Related Behavior

doi: 10.1523/jneurosci.0550-10.2010

Figure Lengend Snippet: Figure1. GeneratingdoublemutantmicedeficientinBmpr1aandBmpr1b.A–C,CoronalsectionsthroughthetelencephalonofcontrolmiceatE11.5,processedwithinsituhybridizationtoshow expression of Bmpr1a (A), Bmpr1b (B), and a third type I BMP receptor gene, Alk2 (C). Bmpr1a is expressed throughout the telencephalic neuroepithelium including the cortical hem; Bmpr1b is expressedmoreselectivelyinthedorsaltelencephalonwithahotspotofexpressionatthehem;Alk2isexpressedbroadlyinthedorsalandventraltelencephalon,butnotinthecorticalhem(C,C). D,BreedingstrategyusedtogeneratemicelackingBmpr1bconstitutivelyandBmpr1aconditionally,theBmpr1afx /;Bmpr1b /;Emx1 /IREScredoublemutantgenotype.E,PCRanalysisofDNA extracted from the tail and telencephalon of E12.5 Bmpr1afx /;Emx1 /IREScre and Bmpr1afx / mice. Primers fx1 and fx4 (Mishina et al., 2002) amplified a 180 bp fragment from Bmpr1afx /; Emx1 /IREScretelencephalon(lane4),indicativeofCre-mediatedrecombinationofBmpr1afx.The180bp“recombined”bandwasnotamplifiedfromtailtissue(lane2)orBmpr1afx /telencephalon (lane 6). Primers fx3 and fx5 amplified a 190 bp fragment from the Bmpr1a constitutive null allele in all three tissue samples (lanes 1, 3, 5). F, G, Coronal sections through the hem region at E12.5 inacontrolmouse(c)andadoublemutant(dm),immunostainedforpSmad1/5/8,transcriptionfactorsactivateddownstreamofBMPsignaling.TheredarrowsindicatepSmad1/5/8-IRcellsinthe controlhem(F)butvirtuallynopSmad1/5/8-IRcellsinthedoublemutanthem(G).Outsidethehem,inthehippocampalprimordium,pSmad1/5/8-IRcellsaredensealongtheventricularsurface (F,G).H,I,Ayoungadultcontrolmouse(H)andlittermatedoublemutant(I).Thedoublemutantisslightlysmallerthanthecontrol;theredarrowsindicatetruncateddigitsandpartiallossoffacial hair(seeResults).Abbreviations:hem,Corticalhem;hp,hippocampus;lge,lateralganglioniceminence;mge,medialganglioniceminence;ncx,neocortex;th,thalamus.Scalebar:(inA)A–C,200 m; C, 100 m; F, G, 50 m.

Article Snippet: For immunohistochemistry, primary antibodies were as follows: antiphospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad8 (Ser426/ 428) rabbit polyclonal antibody (1:50; Cell Signaling Technology), antiphosphohistone H3 (PH3) (Ser10) rabbit polyclonal antibody (1:200; Millipore), anti-calbindin D-28K rabbit polyclonal antibody (1:250; Millipore Bioscience Research Reagents), anti-neuronal nuclei (NeuN) mouse monoclonal antibody (1:10; Millipore), anti-5-bromo-2-deoxyuridine (BrdU) mouse monoclonal antibody (1:75; BD Biosciences) for IHC and anti-BrdU mouse monoclonal antibody (1:50; Serotec) for immunofluorescence, anti-tyrosine hydroxylase (TH) antibody (1:5000; BD Biosciences), anti- -tubulin (Tuj1) (1:500; Covance), anti-Axin2 rabbit polyclonal antibody (1:5000; Abcam), anti-Olig2 rabbit polyclonal antibody (1:1000; Abcam), anti-(cleaved) caspase 3 rabbit polyclonal antibody (1:200; Cell Signaling), anti-Dickkopf1 (DKK1) rabbit polyclonal antibody (1:2000; Santa Cruz Biotechnology), anti-calretinin rabbit polyclonal antibody (1:1000; Millipore), and anti-Acvr1 (ALK2) rabbit polyclonal antibody (1:2000; Novus Biologicals).

Techniques: Expressing, Amplification

Journal: The Journal of endocrinology

Article Title: Activin A and activin receptors in gestational tissue from preeclamptic pregnancies.

doi: 10.1677/joe.0.1710057

Figure Lengend Snippet: Figure 3 Localisation of activin A subunit and receptor Alk2. Panels A and B show activin A staining in healthy term and preeclamptic term placentae respectively. Panel B shows intense staining in syncytiotrophoblast (st) of preeclamptic placenta. Less intense staining was seen in cytotrophoblast (ct), and stroma (s) of healthy and preeclamptic placentae. Fetal membranes from healthy and preeclamptic pregnancies are shown in panels C and D respectively. A staining is present in amniotic epithelium (ep), fibroblast layer (fib), reticular layer (r), chorionic trophoblast (t) and adherent decidua (dec). Receptor Alk2 staining in healthy and preeclamptic placentae is seen mainly in the endothelium (en) of the placental vasculature (panels G and H respectively). The insert in panel G is a serial section with the vasculature stained with the endothelial marker CD34. Alk2 staining in healthy and preeclamptic membranes (panels I and J respectively) is similar to A except that the amniotic epithelium and chorionic trophoblast were mostly negative. Positive and negative control sections (endometrium) for A are shown in panels E and F and panels K and L for Alk2. Scale bars=50 m except panels E, F, K and L where scale bars=25 m.

Article Snippet: Non-specific binding was blocked with 1% gelatine and 0·02% Tween 20 in Tris-buffered saline and blots were incubated with polyclonal goat anti-human activin receptor Alk2, ActRII and ActRIIB antibodies (0·3 μg/ml; R & D Systems, Minneapolis, MN, USA) for 1 h at room temperature.

Techniques: Staining, Marker, Negative Control

Journal: The Journal of endocrinology

Article Title: Activin A and activin receptors in gestational tissue from preeclamptic pregnancies.

doi: 10.1677/joe.0.1710057

Figure Lengend Snippet: Figure 4 Western hybridisation; representative blots with placental, choriodecidual and amnion tissue protein (upper, mid and lower panels respectively) hybridised with activin receptor Alk2, ActRII and ActRIIB antibodies. Each blot contains proteins from T-C, PT-PE, T-PE and endometrium (E, positive control).

Article Snippet: Non-specific binding was blocked with 1% gelatine and 0·02% Tween 20 in Tris-buffered saline and blots were incubated with polyclonal goat anti-human activin receptor Alk2, ActRII and ActRIIB antibodies (0·3 μg/ml; R & D Systems, Minneapolis, MN, USA) for 1 h at room temperature.

Techniques: Western Blot, Hybridization, Positive Control