anti k v 3 4  (Alomone Labs)

Journal: Scientific Reports

Article Title: Kv3.4 regulates cell migration and invasion through TGF-β-induced epithelial–mesenchymal transition in A549 cells

doi: 10.1038/s41598-024-52739-4

Figure Lengend Snippet: Antibody information.

Article Snippet: Kv3.4 , Alomone Labs , APC-019 , 1:500.

Techniques:

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Western blots of wild type (Wt), N220Q, N229Q, and N220Q/N229Q Kv3.1 proteins. Kv3.1 proteins were detected when heterologously expressed in B35 cells (A). Arrows and lines denote the type of N -glycan attached to the Kv3.1 protein. Assignments of the various glycosylated and unglycosylated Kv3.1 proteins were based on immunoband shifts produced by glycosidase treatment. N220Q and N229Q proteins were digested (+) and undigested (−) with neuraminidase (B), PNGase F (C) and Endo H (D). A solid line on image indicates that samples were run on a different blot (B). The numbers adjacent to the Western blots represent the Kaleidoscope markers (in KDa). Similar migration patterns were observed on at least three separate Western blots.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Western Blot, Produced, Migration

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Whole cell currents for glycosylated (A, middle panel; B, top panel) and unglycosylated (A and B, bottom panels) Kv3.1 proteins were elicited from the indicated voltage protocol (A, top panel). Whole cell currents were scaled for inactivating (A) and non-inactivating (B) current types from B35 cells expressing glycosylated and unglycosylated Kv3.1 proteins. Right panels show traces at expanded time scales and grey lines denote currents at +40 and +60 mV. Traces were scaled to show differences in activation kinetics. Conductance-voltage (g/gmax) curves of both inactivating (C) and non-inactivating (D) current types for glycosylated and unglycosylated Kv3.1 channels. Rise times of inactivating (E) and non-inactivating (F) currents types. n represents number of cells.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Expressing, Activation Assay

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Electrophysiological parameters of glycosylated, unglycosylated, and partially glycosylated forms of the Kv3.1 channel in B35 cells.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques:

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: A deactivation voltage protocol (A, left panel) was utilized to obtain deactivation currents for B35 cells expressing the glycosylated (A, right panel) and unglycosylated. Scaled deactivation currents from transfected B35 cells expressing either inactivating (B) or non-inactivating (D) current types. Grey lines denote currents at −30 and −50 mV. Traces were scaled to show differences in deactivation kinetics. Deactivation time constant vs. voltage plot of B35 cells expressing glycosylated and unglycosylated Kv3.1 channels for inactivating (C) and non-inactivating (E) currents types.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Expressing, Transfection

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Whole cell currents were elicited from the shown voltage protocol (A, left panel) for B35 cells expressing glycosylated (A, right panel) and unglycosylated (B) Kv3.1 proteins. Traces were scaled to show differences in inactivation kinetics. Grey lines denote currents at +40 mV.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Currents were elicited by a train of five depolarizing voltage steps to +40 mV once every 525 ms, from a holding potential of −50 mV (A, top panel) for B35 cells expressing glycosylated (A, bottom-left panel) and unglycosylated (A, bottom-right panel) Kv3.1 channels. A bar graph representing the percent of peak current amplitude remaining after the fifth pulse relative to peak current amplitude of initial pulse for the various Kv3.1 channels (B). Asterisks indicate significant differences in mean values at a probability of P <0.01 from that of glycosylated Kv3.1.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Importance of Glycosylation on Function of a Potassium Channel in Neuroblastoma Cells

doi: 10.1371/journal.pone.0019317

Figure Lengend Snippet: Wound width was determined and then normalized at 0, 6, 11 and 23 h for glycosylated and unglycosylated Kv3.1 transfected and non-transfected B35 cells (A). Similar experiments were also performed for glycosylated, and partially glycosylated Kv3.1 glycoproteins (N220Q, and N229Q) transfected B35 cells (B). Data were expressed as the mean +/− SEM. Asterisks indicate significant differences in mean values at a probability of P <0.01 from that of glycosylated Kv3.1. Images were obtained at 0 h, 6 h, 11 h and 23 h of the generated wound for group I: wild type Kv3.1 (row I), N220Q/N229Q (row II) transfected B35 cells and non-transfected B35 cells (row III); and group II: wild type Kv3.1 (row IV), N220Q (row V) and N229Q (row VI) (C). The distance between the two white dashed lines represents wound width for each image. This distance becomes smaller as time increases, representing the rate of cell migration. n represents number of cell wounds. The experiments were conducted on three separate occasions. The solid black line represents a 25 µM scale bar.

Article Snippet: Electrophoresed proteins were transferred to Immobilon-P PVDF membranes (Millipore, Billercia, MA, USA) at 175 mAmps for 90–240 min. Blots were then incubated at room temperature for 20 min in blocking buffer (PBS, 3% BSA with 0.1% Tween 20) followed by incubation for 2 h with polyclonal rabbit anti-Kv3.1, anti-Kv3.3, or anti-Kv3.4 antibodies (Alamone Labs, Jerusalem, Israel) or overnight with mouse anti-Kv3.1 antibody (NeuroMab).

Techniques: Transfection, Generated, Migration