apc101  (Alomone Labs)

Journal: Cancers

Article Title: Inhibition of a Mitochondrial Potassium Channel in Combination with Gemcitabine and Abraxane Drastically Reduces Pancreatic Ductal Adenocarcinoma in an Immunocompetent Orthotopic Murine Model

doi: 10.3390/cancers14112618

Figure Lengend Snippet: Expression of Kv1.3 in human pancreatic cancer tissue. Expression of Kv1.3 in human pancreatic tissue. Panel ( a ), high expression level of Kv1.3 in human pancreatic cancer tissue. ( n = 33 out of 55, scale bar = 50 µm). Panel ( b ), magnification of panel ( a ), scale bar = 20 µm. Panel ( c ), low expression level of Kv1.3 in human pancreatic cancer tissue. ( n = 22 out of 55, scale bar = 50 µm). Panel ( d ), magnification of panel c, scale bar = 20 µm. Panel ( e ), Immunoglobulin G (IgG) control with magnification (Panel ( f )). Panel ( g ) shows a pancreas adenocarcinoma ductule with high expression of Kv1.3 (brown color) and adjacent normal pancreas parenchyma without Kv1.3 expression (black arrow), scale bar = 50 µm. Panel ( h ), magnification of panel ( g ), scale bar = 20 µm.

Article Snippet: Primary antibodies Kv1.3 (1:100, APC-101, Alomone labs, Jerusalem, Israel) and TOM20 (1:100, MABT166, Sigma, Burlington, MA, USA) were incubated overnight at 4 °C.

Techniques: Expressing

Journal: Cancers

Article Title: Mitochondrial Kv1.3 Channels as Target for Treatment of Multiple Myeloma

doi: 10.3390/cancers14081955

Figure Lengend Snippet: Mitochondrial Kv1.3 is expressed in the multiple myeloma cell lines L-363 and RPMI-8226. PAPTP and PCARBTP efficiently kill human multiple myeloma cells. ( a ) Whole cell lysates (left, 50 µg protein/lane) were lysed in RIPA-Buffer, mitochondria from L-363, RPMI-8226 and Jurkat cells (right, 30 µg protein/lane) were enriched as described; both were supplemented with 5× reducing SDS sample buffer, boiled and separated on 8.5% SDS-polyacrylamide gels and blotted on nitrocellulose membranes. Membranes were divided and, after blocking, membranes were incubated 1 h with anti-Kv1.3 or anti-Tim23 primary antibodies, respectively. After extensively washing, blots were incubated with secondary antibodies for 1 h, washed and developed with the Roti–Lumin system. Multiple myeloma cell lines RPMI-8226 and L-363 were treated with increasing concentrations PAPTP ( b ) or PCARBTP ( c ), solvent control (0.1% DMSO), 2 µM staurosporine as a positive control and 2 µM margatoxin as a membrane-impermeable Kv1.3 blocker. After 24 h, cells were stained with Annexin V-APC/7AAD and examined for cell death by flow cytometry. Results are reported as percentages with respect to untreated samples ± SD, ( n = 3 independent experiments, each experiment in triplicate). Graph insert EC50: EC50 values of the indicated compounds in RPMI-8226 and L-363 were calculated by using GraphPad Prism version 9.3 for Windows (GraphPad Software, San Diego, CA, USA).

Article Snippet: Membranes were blocked with 4% BSA/PBS for 1 h and incubated with anti-Kv1.3-antibody (Alomone Labs, Jerusalem, Israel, #APC-101) and anti-Tim23-antibody (BD, Franklin Lakes, NJ, USA, #611223).

Techniques: Blocking Assay, Incubation, Positive Control, Staining, Flow Cytometry, Software

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Comparison of stroke severity in WT versus Kv1.3 −/− mice. (A) Infarct area and deficit score in 16‐week‐old WT ( n = 10) were compared to Kv1.3 −/− ( n = 14) male mice ( p

Article Snippet: On day 8, animals were sacrificed and 14‐µ m thick cryosections were stained with a polyclonal rabbit anti‐Kv1.3 antibody (Alomone, APC‐101, 1:1000) followed by an Alexa Fluor® 546‐conjugated secondary antibody (1:1000, Life Technologies).

Techniques: Mouse Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: The Kv1.3 blocker PAP‐1 reduces infarction and improves neurological deficit in young and old WT female mice. (A) Infarct area and deficit score in vehicle ( n = 11) were compared to PAP‐1 (40 mg/kg; n = 15) treated 16‐week‐old female mice ( p = 0.007 for infarct, p = 0.016 for NES). (B) Infarct area and deficit score in vehicle ( n = 9) were compared to PAP‐1 (40 mg/kg; n = 9) treated 80‐week‐old female mice ( p = 0.032 for infarct, p = 0.003 for NES). Data are shown as whisker plots with data overlay. The boxes show mean ± SEM and the whiskers show confidence intervals.

Article Snippet: On day 8, animals were sacrificed and 14‐µ m thick cryosections were stained with a polyclonal rabbit anti‐Kv1.3 antibody (Alomone, APC‐101, 1:1000) followed by an Alexa Fluor® 546‐conjugated secondary antibody (1:1000, Life Technologies).

Techniques: Mouse Assay, Whisker Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Kv1.3 is expressed on microglia/macrophages and T cells in the infarct of an 80‐week‐old female mouse. (A) Sample immunofluorescence staining of 5‐ µ m thick paraffin sections from the 6‐mm slice of an 80‐week‐old female mouse on day 8 after tMCAO. (A) Kv1.3 staining colocalizes to Iba‐1 + cells. (B) Kv1.3 staining on smaller, CD3 + T cells (white arrowheads).

Article Snippet: On day 8, animals were sacrificed and 14‐µ m thick cryosections were stained with a polyclonal rabbit anti‐Kv1.3 antibody (Alomone, APC‐101, 1:1000) followed by an Alexa Fluor® 546‐conjugated secondary antibody (1:1000, Life Technologies).

Techniques: Immunofluorescence, Staining

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: PAP‐1 treatment and Kv1.3 deletion reduce Iba‐1 and CD3 staining in 80‐week‐old female mice. Serial paraffin sections (5 µ m thick) cut at 4 mm and the 6 mm from 80‐week‐old female mice were stained for Iba‐1 and CD3. (A) Percentage of Iba‐1‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.120 for 4‐mm section; p = 0.014 for 6‐mm section), and Kv1.3 −/− ( n = 18, p = 0.011 for 4‐mm section; p = 0.007 for 6‐mm section). (B) Percentage of CD3‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.049 for 4‐mm section; p = 0.027 for 6‐mm section), and Kv1.3 −/− ( n = 18, p

Article Snippet: On day 8, animals were sacrificed and 14‐µ m thick cryosections were stained with a polyclonal rabbit anti‐Kv1.3 antibody (Alomone, APC‐101, 1:1000) followed by an Alexa Fluor® 546‐conjugated secondary antibody (1:1000, Life Technologies).

Techniques: Staining, Mouse Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Comparison of stroke severity in WT versus Kv1.3 −/− mice. (A) Infarct area and deficit score in 16‐week‐old WT ( n = 10) were compared to Kv1.3 −/− ( n = 14) male mice ( p

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Mouse Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: The Kv1.3 blocker PAP‐1 reduces infarction and improves neurological deficit in young and old WT female mice. (A) Infarct area and deficit score in vehicle ( n = 11) were compared to PAP‐1 (40 mg/kg; n = 15) treated 16‐week‐old female mice ( p = 0.007 for infarct, p = 0.016 for NES). (B) Infarct area and deficit score in vehicle ( n = 9) were compared to PAP‐1 (40 mg/kg; n = 9) treated 80‐week‐old female mice ( p = 0.032 for infarct, p = 0.003 for NES). Data are shown as whisker plots with data overlay. The boxes show mean ± SEM and the whiskers show confidence intervals.

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Mouse Assay, Whisker Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Kv1.3 is expressed on microglia/macrophages and T cells in the infarct of an 80‐week‐old female mouse. (A) Sample immunofluorescence staining of 5‐ µ m thick paraffin sections from the 6‐mm slice of an 80‐week‐old female mouse on day 8 after tMCAO. (A) Kv1.3 staining colocalizes to Iba‐1 + cells. (B) Kv1.3 staining on smaller, CD3 + T cells (white arrowheads).

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Immunofluorescence, Staining

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: PAP‐1 treatment and Kv1.3 deletion reduce Iba‐1 and CD3 staining in 80‐week‐old female mice. Serial paraffin sections (5 µ m thick) cut at 4 mm and the 6 mm from 80‐week‐old female mice were stained for Iba‐1 and CD3. (A) Percentage of Iba‐1‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.120 for 4‐mm section; p = 0.014 for 6‐mm section), and Kv1.3 −/− ( n = 18, p = 0.011 for 4‐mm section; p = 0.007 for 6‐mm section). (B) Percentage of CD3‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.049 for 4‐mm section; p = 0.027 for 6‐mm section), and Kv1.3 −/− ( n = 18, p

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Staining, Mouse Assay