Journal:

Article Title: Role of transverse bands in maintaining paranodal structure and axolemmal domain organization in myelinated nerve fibers: effect on longevity in dysmyelinated mutant mice

doi: 10.1002/cne.22367

Figure Lengend Snippet: Details of antibodies used for immunostaining and western blots.

Article Snippet: Kv1.2 , Fusion protein amino acids 428–499(cytoplasmic C-terminus) of rat Kv1.2 (accession number {"type":"entrez-protein","attrs":{"text":"P63142","term_id":"52000923","term_text":"P63142"}} P63142 ), Epitope mapped to within amino acids 463–480 , Neuromab Balb/C mouse monoclonal 75-008 , 1:200.

Techniques: Immunostaining, Western Blot, Staining, Sequencing, Bioprocessing

Journal: The Journal of Neuroscience

Article Title: Tonotopic Specialization of Auditory Coincidence Detection in Nucleus Laminaris of the Chick

doi: 10.1523/JNEUROSCI.4428-04.2005

Figure Lengend Snippet: Distribution of Kv1.2 channel protein along the tonotopic axis. A, Transverse slice stained with the Kv1.2 antibody. Boxes show the regions of NL magnified in B-D. The left panel in A and bottom panels in E-G show the relative fluorescence intensity of individual NL neurons (see Materials and Methods). Numbers on the abscissa indicate the sector of NL. B-D, Immunoreactivity is strong in the middle region (C) and decreases in both the rostromedial (B) and caudolateral (D) region (p < 0.01 by Mann-Whitney U test among the three CF regions in a slice). E-G, Coronal slices stained with the antibody. R, Rostral; M, medial; D, dorsal. The immunoreactivity increases from sector 2 to sector 3 in slice E (high CF) but decreases in slice F from sectors 7-8 (middle CF) to sector 9 (low CF) and decreases in slice G toward the lower-CF sector (p < 0.01 by Mann-Whitney U test among sectors in each slice). H, This gradient is developed along the tonotopic axis, which is directed from the rostromedial high CF to the caudolateral low CF. Each slice is illustrated by a dotted line in H. I, Relative fluorescence intensity in each sector. Data from four chicks were averaged, and comparisons were made among sectors within the same slices (see Materials and Methods). #Statistical significance for p < 0.05.

Article Snippet: Preadsorption of the antibody with the Kv1.2-glutathione S -transferase (GST) fusion protein (3 μg/ml; Alomone Labs) blocked the staining in the chicken brain sections.

Techniques: Staining, Fluorescence, MANN-WHITNEY

Journal:

Article Title: Potassium channels Kv1.1, Kv1.2 and Kv1.6 influence excitability of rat visceral sensory neurons

doi: 10.1113/jphysiol.2001.018333

Figure Lengend Snippet: A, Kv1.1, Kv1.2 and Kv1.6 channel mRNAs are expressed in nodose ganglia. PCR products, resulting from the amplification of first-strand cDNA prepared with (+) or without (-) reverse transcriptase (RT) from nodose ganglia or brain poly A+ RNA with Kv1.1, Kv1.2 or Kv1.6 specific oligomers, were separated by electrophoresis and transferred to nylon membranes. After Southern hybridization with 32P-labelled specific internal oligomers (see Methods), the autoradiogram showed positive signals for all three channels from nodose and rat brain in the (+)-RT lanes with no signals in the control (-)-RT lanes. B, Northern blot analysis of Kvβ1.2 expression in adult rat nodose ganglia and brain. RNA size markers are indicated on the left. As shown, brain exhibited a single band of ∼5 kb that hybridized with a Kvβ1.2 specific riboprobe, a band that is not present in nodose ganglia. C, RT-PCR analysis of the expression of Kvβ1.1, Kvβ1.3, Kvβ2 and Kvβ3 subunits in rat brain and nodose ganglia. As a control, first-strand cDNA reactions were performed either with (+) or without (-) RT. The oligonucleotide probes amplify a cDNA of 323 bp for Kvβ1.1, 182 bp for Kvβ1.3, 524 bp for Kvβ2 and 515 bp for Kvβ3.

Article Snippet: Two Kv1.2 antibodies were used: a rabbit polyclonal (Alomone Labs) raised against a GST fusion protein coupled to a C-terminal portion of rat Kv1.2 (amino acids 417-498) and a mouse monoclonal (Upstate Biotechnology) raised against a GST fusion protein coupled to amino acids 428-499 of rat Kv1.2.

Techniques: Amplification, Electrophoresis, Hybridization, Northern Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: Potassium channels Kv1.1, Kv1.2 and Kv1.6 influence excitability of rat visceral sensory neurons

doi: 10.1113/jphysiol.2001.018333

Figure Lengend Snippet: Western blots of channel expression in nodose ganglia (A, B and C), brain lysates (A and B) and brain crude membrane fraction (C) probed with monoclonal anti-Kv1.1 (A), monoclonal anti-Kv1.2 (B) and polyclonal anti-Kv1.6 (C) (50 μg protein per lane). Immunoreactive bands were visualized with ECL-Plus (Amersham Pharmacia Biotech). Molecular weight markers (kDa) are indicated on the left.

Article Snippet: Two Kv1.2 antibodies were used: a rabbit polyclonal (Alomone Labs) raised against a GST fusion protein coupled to a C-terminal portion of rat Kv1.2 (amino acids 417-498) and a mouse monoclonal (Upstate Biotechnology) raised against a GST fusion protein coupled to amino acids 428-499 of rat Kv1.2.

Techniques: Western Blot, Expressing, Molecular Weight

Journal:

Article Title: Potassium channels Kv1.1, Kv1.2 and Kv1.6 influence excitability of rat visceral sensory neurons

doi: 10.1113/jphysiol.2001.018333

Figure Lengend Snippet: A, left panels show confocal single slice images through isolated neonatal nodose neurons labelled with Kv1.1 (top), Kv1.2 (middle) and Kv1.6 (bottom). The arrows indicate some regions where patches of immunoreactivity appear at the cell membrane. Differential interference contrast (DIC) images of the cells appear in insets in each panel. Right panels show preabsorption controls with the immunizing fusion protein for the three antibodies. B, expression of Kv1.1, Kv1.2 and Kv1.6 in superior laryngeal sensory neurons is shown with conventional fluorescent microscopy of 6 μm sections through the nodose ganglion. The Cm-DiI label was co-localized with the potassium channel antibodies (green) to give a yellow-orange colour. These images were digitally recorded as single labels and then superimposed. These figures also illustrate a wide distribution in the level of immunoreactivity among the neurons. The arrows in the upper panel show that Kv1.1 immunoreactivity is also found in pericytes surrounding the neurons. The section in the middle panel contains a group of nerve fibres. The arrowhead indicates Kv1.2 immunoreactivity in the juxta-paranodal regions of a myelinated fibre and the double-headed arrow indicates immunoreactivity distributed along a finer fibre. In all panels the asterisks identify neurons containing DiI.

Article Snippet: Two Kv1.2 antibodies were used: a rabbit polyclonal (Alomone Labs) raised against a GST fusion protein coupled to a C-terminal portion of rat Kv1.2 (amino acids 417-498) and a mouse monoclonal (Upstate Biotechnology) raised against a GST fusion protein coupled to amino acids 428-499 of rat Kv1.2.

Techniques: Isolation, Expressing, Microscopy