goat polyclonal anti tag1  (R&D Systems)

Journal: Science Advances

Article Title: A global gene regulatory program and its region-specific regulator partition neurons into commissural and ipsilateral projection types

doi: 10.1126/sciadv.adk2149

Figure Lengend Snippet: ( A ) The schematic shows the PN formation in an E16.5 hindbrain. Red dashed lines mark the rostral (r) and caudal (c) span of the sections shown on the right. PN is visualized by Barhl1 IHC. PN neurons form a nucleus adjacent to the ventral midline (arrows) ( n = 3) in the control but were laterally positioned (arrowheads) indicating failure to approach the ventral midline in the mutants ( n = 3). ( B to D ) Coronal sections from the spinal cord (B), hindbrain (C), and midbrain (D) at E11.5 were subjected to Tag1 and NF double IHC with the Tag1 labeling the commissural axons and the NF signals depicting the general axonal patterns. The DAPI counterstain indicates the overall cytoarchitecture. Comparisons were made between the control genotype ( Nhlh1 +/+ Nhlh2 +/m ) ( n = 3) and the double mutant ( Nhlh1 m/m Nhlh2 m/m ) ( n = 3). The Tag1 and NF merged images in the bottom panel are high-magnification images of the ventral commissure regions. The double mutant showed a complete lack of ventral commissures (hollow arrows), in comparison to the control (filled arrows). Scale bars, 200 μm [(A) to (D)].

Article Snippet: The primary antibodies used were rabbit anti-Barhl1 polyclonal antibody (Atlas Antibodies; HPA004809, Sigma-Aldrich; 1:500), goat anti-Robo3 polyclonal antibody (R&D Systems, AF3076; 1:200), goat anti-Tag1 polyclonal antibody (R&D Systems, AF4439; 1:500), mouse anti–NF-160kD monoclonal antibody (clone RMO-270; Thermo Fisher Scientific, 13-0700; 1:500), rat anti-L1CAM monoclonal antibody (clone 324; Merck Millipore, MAB5272; 1:400), goat anti-DCC polyclonal antibody (Santa Cruz Biotechnology, sc-6535; 1:200), rabbit anti-FoxP2 polyclonal antibody (Abcam, ab16046; 1:1000), mouse anti-Brn3a monoclonal antibody (clone 5A3.2; Merck Millipore, MAB1585; 1;200), rabbit anti-Lim1/Lhx1 antibody (Abcam, ab229474; 1:250), rabbit anti-Lhx2 (Invitrogen, PA5-78287; 1:250), rat anti-HA (Roche; clone3F10; 1:100), guinea pig anti-Isl1 antibody (a gift from Y. Tanabe, Kyoto University, Japan), and chick anti-GFP polyclonal antibody (Abcam, ab13970; 1:1500).

Techniques: Control, Labeling, Mutagenesis, Comparison

Journal: The Journal of Clinical Investigation

Article Title: In vivo visualization and molecular targeting of the cardiac conduction system

doi: 10.1172/JCI156955

Figure Lengend Snippet: ( A ) Antibody-dye conjugate (mCntn2-800) consists of a near-infrared (NIR) dye conjugated to an antibody against the CCS-specific surface marker Cntn2 (contactin 2). ( B ) Experimental work flow. ( C ) Whole-body biodistribution of other tissue types, showing expected clearance within the liver, bladder, and kidneys and notable absence from the brain. ( D ) Whole mouse heart ( n = 3) from a wild-type (WT) mouse injected 3 days prior with mCntn2-800, in posterior-anterior (PA) and right lateral (RL) views. Atria are outlined in white and cardiac chambers are listed. LA, left atrium; LV, left ventricle; RA, right atrium; RV, right ventricle. Top: Brightfield. Bottom: NIR signal demonstrating labeling of the CCS (blue→red = lowest→highest signal). Mean signal to background ratio (SBR) is indicated. ( E ) Measured intervals (in ms) from sedated surface electrocardiograms (ECGs) including PR, QRS, QTc, and RR in WT mice prior to (day 0 = baseline, n = 12) and daily (day 1 n = 3, day 2 n = 9; after injection) following a single tail vein injection of mCntn2-800. Intervals (mean ± SD) on a given day after injection were compared to each mouse’s preinjection control baseline (day 0) using 1-way ANOVA with Tukey’s post hoc test. ( F – H ) Heart sections from a WT mouse injected 2 days prior with mCntn2-800. CCS components labeled with mCntn2-800 (purple) and costained with antibodies targeting known markers of the CCS, including anti-Hcn4 (red, SAN and His) or anti-Cx40 (green, PF). DAPI (blue, nuclei). LBB/RBB, left and right bundle branches; His, His bundle; PF, Purkinje fibers; SAN, sinoatrial node; VM, ventricular myocardium. Scale bars: 10 mm ( C ), 5 mm ( D ), and 100 μm ( F – H ).

Article Snippet: Described antibody-dye conjugates consist of commercially acquired anti-Cntn2 goat polyclonal antibody (AF4439) and anti-NPTN goat polyclonal antibody (AF5360) (both R&D Systems) that have been covalently conjugated to a benign, NIR dye (IRDye800CW NHS ester; LI-COR Biosciences, 929-70020) using company specifications.

Techniques: Marker, Injection, Labeling, Control

Journal: The Journal of Clinical Investigation

Article Title: In vivo visualization and molecular targeting of the cardiac conduction system

doi: 10.1172/JCI156955

Figure Lengend Snippet: ( A ) Experimental workflow: wild-type mice ( n = 3) received a tail vein injection of mCntn2-800 and after 48 hours were sedated and received a sternotomy and cardiac incisions including a right atriotomy and right ventriculotomy to simulate a surgical scenario. Live imaging of the heart with a FLARE Intraoperative Near-Infrared (NIR) Fluorescence Imaging System. ( B and C ) Heart with visible sinoatrial node (SAN), atrioventricular node/His bundle (AVN/His), and Purkinje fiber (PF) network. Left: Color image of ex vivo heart. Right: mCntn2-800 NIR signal (green). Middle: Merged image of color image and NIR (green) signal. ( D ) Heart sections from the same heart, demonstrating mCntn2-800 signal (cyan) labeling the SAN, as costained with anti-Cntn2 (red) immunostaining. ( E ) Magnified region in SAN indicated by the white box in D . Right atrial (RA) tissue labeled with anti-Cx43 (purple). DAPI (blue, nuclei). IVS, interventricular septum; RV, right ventricle; SAN, sinoatrial node; SVC, superior vena cava. Scale bars: 5 mm ( B and C ), 300 μm ( D ), and 100 μm ( E ).

Article Snippet: Described antibody-dye conjugates consist of commercially acquired anti-Cntn2 goat polyclonal antibody (AF4439) and anti-NPTN goat polyclonal antibody (AF5360) (both R&D Systems) that have been covalently conjugated to a benign, NIR dye (IRDye800CW NHS ester; LI-COR Biosciences, 929-70020) using company specifications.

Techniques: Injection, Imaging, Fluorescence, Ex Vivo, Labeling, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: In vivo visualization and molecular targeting of the cardiac conduction system

doi: 10.1172/JCI156955

Figure Lengend Snippet: ( A ) Human anti-CNTN2 Fab antibody was biotinylated and conjugated to streptavidin-linked saporin, a cell toxin (hCNTN2-Sap). ( B ) Wild-type mice received a single tail vein injection of either hCNTN2-Sap (100 μg) ( n = 6) or Control-Sap (100 μg nonspecific human IgG similarly conjugated to saporin) ( n = 6). Mice received electrocardiograms (ECGs) on day 0 (baseline) and daily following injection with control or hCNTN2-Sap. On day 2, hearts were harvested, fixed, and immunostained. ( C ) Representative ECG tracings ( n = 6 per condition). ( D ) By day 2, mice injected with hCNTN2-Sap demonstrated marked conduction abnormalities, including prolonged PR, QRS, and RR intervals as compared with mice injected with Control-Sap (mean intervals in ms ± SD). Gray bar, QTc interval corrected for QRS intervals. Statistical analyses using 2-way ANOVA with Tukey’s post hoc test. ( E ) Consistent with targeted cell death, immunofluorescence of the CCS (red) showed subtotal loss of CCS cells as shown within the His bundle (His), right and left bundle branches (RBB/LBB) as stained by anti-Cntn2. His, His bundle; IVS, interventricular septum; LBB, left bundle branch; RBB, right bundle branch. Scale bar: 50 μm.

Article Snippet: Described antibody-dye conjugates consist of commercially acquired anti-Cntn2 goat polyclonal antibody (AF4439) and anti-NPTN goat polyclonal antibody (AF5360) (both R&D Systems) that have been covalently conjugated to a benign, NIR dye (IRDye800CW NHS ester; LI-COR Biosciences, 929-70020) using company specifications.

Techniques: Injection, Control, Immunofluorescence, Staining