Connexin Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina"
Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina
Journal: The Journal of comparative neurology
Figure Legend Snippet: Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.
Techniques Used: Labeling, Incubation, Produced, Expressing
2) Product Images from "Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36"
Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36
Journal: Cell Death & Disease
Figure Legend Snippet: Brainstem Cx36 expression decreases because of nicotine or carbenoxolone application. ( a ) Example of western immunoblotting (top) showing the expression of Cx36 in brainstems incubated for 4 h in Krebs, or treated with TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. Histograms (bottom) quantifying the significantly decreased Cx36 levels after treatment with nicotine + TBOA (n, N =9) or carbenoxolone + TBOA (n, N =5; Kruskal–Wallis one-way ANOVA: P =0.015 among groups). ( b and c ) Example of western blot (top) showing Cx36 expression in membrane ( b ) or the cytoplasmic ( c ) fractions of samples treated as described above. Histograms illustrate how nicotine alone or co-applied induces a significant decrease of Cx36 expression in the membrane pool ( b , bottom; Kruskal–Wallis one-way ANOVA: P =0.069 among groups; Mann–Whitney test: * P =0.016 for sham versus nicotine + TBOA and ** P =0.008 for sham versus nicotine; n, N =5) and an increase in the cytoplasmic one ( c , bottom; Kruskal–Wallis one-way ANOVA: P =0.014 among groups; Tukey test: * P
Techniques Used: Expressing, Western Blot, Incubation, MANN-WHITNEY
Figure Legend Snippet: Cx36 mRNA levels and Hsp70 expression. ( a ) Plot illustrating unchanged values of Cx36 mRNA levels among different treatments. Fractional Cx36 expression (AU): 1±0.0, sham; 0.9±0.2, TBOA; 1±0.3, nicotine + TBOA; 1.3±0.2, nicotine (n, N =6). ( b ) Example of Hsp70 immunoblotting. ( c ) Histogram quantifying the significant increase of Hsp70 expression after nicotine + TBOA treatment (Kruskal–Wallis one-way ANOVA: P =0.027 among groups; Holm–Sidak method: * P
Techniques Used: Expressing
Figure Legend Snippet: Idealized diagram to account for the toxic effects by TBOA and their antagonism by nicotine on HMs. ( a ) In the presence of TBOA, glutamate (filled red circles) released from presynaptic terminals acts on glutamatergic receptors to depolarize HMs. TBOA-mediated inhibition of glutamate uptake by astrocytes leads to intensification of glutamate effects with build-up of intracellular free Ca 2+ , which acts on mitochondria and favors production of ROS, release of AIF and DNA damage. This process is greatly amplified by the bidirectional (red arrows) operation of gap junctions (Cx36; filled blue cylinders) that spread and recruit HMs into a hyperexcitable state with deleterious consequences on cell survival. ( b ) In the presence of nicotine, the neurotoxic effect of TBOA is largely attenuated. Nicotine exerts multiple effects via nAChRs located on presynaptical terminals (with consequent decrease in glutamate release, and increase in the inhibitory response), HM membrane and even mitochondria. Through these effects, nicotine strongly decreases the motoneuron coupling via gap junctions through redistribution of Cx36 subunits, therefore disjoining motoneurons from their collective excitatory behavior. Our hypothesis is that this process is accompanied by inhibition of Cx36 activity, reduction in production of ROS and facilitation of the effect of Hsp70 to sequester AIF and prevent its nuclear migration. For references, see text
Techniques Used: Inhibition, Amplification, Activity Assay, Migration
Figure Legend Snippet: Nicotine and carbenoxolone induce significant reduction in Cx36 expression. ( a ) Example of HMs after 4-h incubation in Krebs (sham), TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. HMs were labeled with SMI 32 (green pseudocolor; left column), as motoneuronal marker, or Cx36 (red pseudocolor; middle column). The right column shows merged images where DAPI (blue pseudocolor) is used to visualize nuclei. ( b ) Histograms quantify the Cx36 signal decrease after 4h of nicotine + TBOA (Kruskal–Wallis one-way ANOVA: P =0.005 among groups; Dunn’s method: * P
Techniques Used: Expressing, Incubation, Labeling, Marker