anti kv2 2  (Alomone Labs)

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Regulation of Kv2.2 by pyruvate-isocitrate cycling and regulation of GSIS by Kv2.2. 832/13 cells were treated with siControl ( siCont ), siCIC, or siICDc for 72 h, and total RNA was isolated for qRT-PCR analyses of CIC, ICDc, Kv2.1, and Kv2.2 expression

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Isolation, Quantitative RT-PCR, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Kv2.2 is expressed in α-, β- and δ-cells of the intact rodent islet. A–D , dissociated mouse islet cells were sorted based on expression of fluorescent proteins and tested for Kv2.2 expression by qRT-PCR analyses. A , transgenic

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Expressing, Quantitative RT-PCR, Transgenic Assay

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Kv2.2 overexpression rescues the inhibitory effects of siICDc on GSIS in primary islets. Freshly isolated rat islets were transduced with an equal dose of Ad-siControl ( siCont ) or Ad-siICDc ( siICDc ) and subsequently incubated for a total of 72 h. A , for

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Over Expression, Isolation, Transduction, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Inhibitory effects of siICDc on GSIS are rescued by Kv2.2 overexpression in 832/13 cells. Immunoblot analyses of membrane protein fractions isolated from untreated (−) and from AdCMV-hKv2.2 (Kv2.2 OE)-treated 832/13 cells are shown. The AdCMV-hKv2.2

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Over Expression, Isolation

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Interactions of Kv2.2 and Kv2.1 regulate outward K + current in β-cells. 832/13 cells were transfected with siControl ( siCont ), siICDc, siKv2.1, or siKv2.2 duplexes in combination with a plasmid encoding GFP and cultured for 72 h prior to patch

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Transfection, Plasmid Preparation, Cell Culture

Journal: The Journal of Biological Chemistry

Article Title: Control of Voltage-gated Potassium Channel Kv2.2 Expression by Pyruvate-Isocitrate Cycling Regulates Glucose-stimulated Insulin Secretion *

doi: 10.1074/jbc.M113.491654

Figure Lengend Snippet: Kv2.2 mRNA expression is reduced by chronic exposure to elevated fatty acids. 832/13 cells were cultured for a total of 3 days and exposed to a mixture of 1 m m O/P complexed to BSA in a 2:1 ratio or to BSA alone for the final 24 or 48 h as indicated.

Article Snippet: These results were confirmed by blotting with an anti-Kv2.2 antibody ( B ).

Techniques: Expressing, Cell Culture

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Reciprocal expression of Kv2.1 and Kv2.2 in the rat brain. A–C: Expression pattern of Kv2.1 ( A ) and Kv2.2 ( B ) in the rat brain. Dorsal to ventral view of rat sagittal brain sections immunostained with anti-Kv2.2 and K89 anti-Kv2.1 antibodies. The cortex (CTX), the striatum (STR) and the basal forebrain (BF) are shown. Mouse Kv2.2 in situ hybridiztion results from the Allen Brain Institute confirms Kv2.2 immunohistochemical data ( C ). Scale bar, 1 mm. D–G: Differential expression pattern of Kv2.1 and Kv2.2 in the cortex and the BF. Kv2.1 is highly expressed in neurons in the cortex and shows decreased expression levels in the BF. Kv2.2 is highly expressed in the BF and is only expressed in a subset of pyramidal neurons mainly in layers I/II and V at low expression levels. Scale bar, 50 μm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, In Situ, Immunohistochemistry

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Kv2.2 expression defines a novel subpopulation of GABAergic neurons in the BF. Stereological analysis of three consecutive coronal brain sections spanning 120 μm of the MCPO/HDB (each 40 μm thick) from GAD67-GFP knock-in mice (n=3). These sections were immunostained with anti-Kv2.2, anti-GFP, and anti-NeuN antibodies. Labeled neurons were counted per section/per animal and averaged among animals.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Knock-In, Mouse Assay, Labeling

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: The expression of Kv2.2 in non-cholinergic neurons in the BF. Kv2.2-expressing neurons and cholinergic neurons are distinct sub-populations in the BF. Rat coronal sections were double immunostained with anti-Kv2.2 and anti-ChAT antibodies. Scale bar, 50 μm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Specificity of the anti-Kv2.2 antibody. A: Rat brain membrane fraction was solubilized and subjected to Western blotting with (right lane) or without (left lane) the pre-absorption with the antigen peptide (1 μg/ml peptide to 0.1 μg/ml antibody). B: Brain lysates from wild-type, Kv2.1-deficient, and Kv2.2-deficient mice were subjected to Western blotting using K89 (top) or anti-Kv2.2 antibody (bottom). Numbers on the left denote the sizes of molecular weight markers in kDa. C: Immunostaining of HEK293 cells doubly transfected with Kv2.1 and Kv2.2. The arrows indicate cells solely stained either by K89 or Kv2.2 antibodies, and the arrowhead indicates a double-labeled cell. Non-transfected cells in the field were not labeled. The specificity was also verified in cells transfected with individual Kv2 subunit (not shown). D–F: Rat sagittal sections (lateral 2.62 mm) are stained with K89 anti-Kv2.1 and anti-Kv2.2 antibodies. The ventral surface of the brain between the nucleus of lateral olfactory tract (LOT) and the olfactory tubercle (Tu) is shown. AC, anterior commissure. Inset in F shows the rat brain atlas of the respective sagittal section. GP, globus pallidus; CPu, caudative putamen; EA, extended amygdala; VP, ventral pallidum. Scale bar, 1 mm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Western Blot, Mouse Assay, Molecular Weight, Immunostaining, Transfection, Staining, Labeling

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Abundant expression of Kv2.2 in the BF of the rat. A–C: Overlapping patterns of immunolabeling with rabbit anti-Kv2.2C (Rabbit) and K37 anti-Kv2.2N (Mouse) antibodies. Scale bar, 100 μm. D: Line scanning analysis of a 450 μm segment in the fluorescence image in F. Green, rabbit Kv2.2 antibody; magenta, mouse Kv2.2 antibody. Overlap of signals is indicated in yellow. E–F: Kv2.2 immunostaining of heterozygous and homozygoous Kv2.2-deficient mice (Kv2.2 +/− and Kv2.2 −/− , respectively. Brain sections were stained using rabbit Kv2.2 antibody. Scale bar, 100 μm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Immunolabeling, Fluorescence, Immunostaining, Mouse Assay, Staining

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Expression of Kv2.2 in the neurons of the magnocellular preoptic nucleus (MCPO) and the horizontal band of Broca (HDB). A–C: Expression of Kv2.2 in the NeuN-positive neuronal population in the basal forebrain. Scale bar, 50 μm. D–E: Nickel enhanced 3-3′diaminobenzidine (DAB) immunostaining was used to determine specific localization of Kv2.2-expressing neurons. Rat coronal sections (8.9 and 9.2mm anterior to the interaural line) were immunostained with the anti-Kv2.2 antibody. Anatomical landmarks such as the anterior commissure (indicated by asterisk) were used to locate the expression of Kv2.2 in the MCPO/HDB nuclei.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Immunostaining

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Selective expression of Kv2.2 in GABAergic neurons in the BF of mouse. A–C: Selective expression of GFP in GABAergic neurons in the cortrex of the GAD67-GFP knock-in mice. Coronal sections from GAD67-GFP knock-in mice were double immunostained with N86 anti-GFP and anti-GABA antibodies. Scale bar, 50 μm. D–F: GFP expressing neurons and ChAT positive neurons of the BF are distinct populations in the BF of the GAD67-GFP knock-in mice. Coronal sections from GAD67-GFP knock-in mice were double immunostained with N86 anti-GFP and anti-ChAT antibodies. Scale bar, 100 μm. G–I: Co-expression of Kv2.2 and GFP in the BF neurons of GAD67-GFP knock-in mice. Coronal sections from GAD67-GFP knock-in mice were double immunostained with anti-Kv2.2 and N86 anti-GFP antibodies. Scale bar, 20 μm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Knock-In, Mouse Assay

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Enriched expression of Kv2.2 in non-cholinergic neurons in the MCPO/HDB of the mouse brain. A: Nickel enhanced DAB immunostaining of Kv2.2-immunoreactive neurons in the mouse brain. A coronal section (3.94 mm anterior to the interaural line) was immunostained with the anti-Kv2.2 antibody. B: Corresponding mouse brain atlas to A. VLPO, ventrolateral preoptic nucleus; LPO, lateral preoptic nucleus; VP, ventral pallidum; AC, anterior commissure; SIB, substantia innominata basal; Tu, olfactory tubercle. C–E: Confirmation of the enrichment of Kv2.2 in the MCPO in immunofluorescence staining. Mouse coronal sections were double immunostained with K89 anti-Kv2.1 and anti-Kv2.2 antibodies. F–H: Reciprocal expression of Kv2.2 and ChAT in the MCPO of mouse. Mouse coronal sections were double immunostained with anti-Kv2.2 and anti-ChAT antibodies. Scale bars, 100 μm.

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Immunostaining, Immunofluorescence, Staining

Journal: The Journal of comparative neurology

Article Title: Immunolocalization of the Voltage-gated Potassium Channel Kv2.2 in GABAergic Neurons in the Basal Forebrain of Rats and Mice

doi: 10.1002/cne.22457

Figure Lengend Snippet: Reciprocal expression of Kv2.1 and Kv2.2 revealed by quantitative Western blotting. A: Micropunch tissue samples (0.5–1 mm) from the cortex were prepared and processed for western blot analysis using K89 anti-Kv2.1 and rabbit anti-Kv2.2 antibodies. Semi-linear relationships were obtained between immunoreactivity and the amount of samples for each of the antibodies. B: Micropunch tissues from the MCPO and the cortex was subjected to Western blotting using K89 anti-Kv2.1, anti-Kv2.2 and anti-NeuN antibodies. Numbers on the both ends denote the sizes of molecular weight markers in kDa. C: The relative levels of each Kv2 isotypes in the MCPO. Optical densities of immunoreactive bands in the MCPO samples were normalized to those of the cortex samples. Values are expressed as means ± SEM. *P

Article Snippet: For DAB staining, sections were incubated with rabbit anti-Kv2.2 antibody (0.3 μg/ml, Alomone Labs, Jerusalem, Israel) overnight, washed, and probed with a biotinylated anti-rabbit secondary antibody (1:250, Vector Labs, Burlingame, CA).

Techniques: Expressing, Western Blot, Molecular Weight