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Apc 087

Manufactured by Alomone
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APC-087 is a labeling reagent for flow cytometry. It is designed to conjugate with specific antibodies and be used in the analysis of cellular samples.

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Lab products found in correlation

Alexa fluor 488 conjugated donkey anti rabbit, by Thermo Fisher Scientific (2 mentions) P6154, by Avantor (2 mentions) Nbp1 78446, by Alomone (2 mentions) Nucblue, by Thermo Fisher Scientific (2 mentions) Alexa fluor 568 phalloidin, by Thermo Fisher Scientific (2 mentions) Dylight 488 conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Ab2413, by Abcam (1 mentions) Ab34710, by Abcam (1 mentions) Axio observer z1, by Zeiss (1 mentions) Cy3 conjugated anti mouse igg, by Jackson ImmunoResearch (1 mentions) Acc 034, by Alomone (1 mentions) A7030, by Merck Group (1 mentions) Optical microscope, by Olympus (1 mentions)

5 protocols using apc 087

1

Immunostaining of Mouse and Human PSCs

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Mouse and human PSCs were washed with PBS (pH 7.4) and fixed with 4% paraformaldehyde for 10 minutes at room temperature. The fixed cells were treated with 0.1% Triton X-100 (13 (link)). Cells were immunostained with a rabbit anti–TRPV4 antiserum (Alomone, ACC-034, 1:250; ref. 66 (link)), rabbit anti–Piezo1 antiserum (Alomone, APC-087, 1:300; ref. 11 (link)), rabbit anti-fibronectin antibody (Abcam, ab2413), rabbit anti–collagen type I antibody (Abcam, ab34710), or chicken anti-GFAP antibody (Abcam, 4674) for mouse PSCs or rabbit anti-GFAP (Cell Signaling Technology,12389) for human PSCs overnight at 2°C–8°C. Secondary antibodies included DyLight 488–conjugated anti–chicken IgG (Jackson ImmunoResearch, 703-546-155), DyLight 488–conjugated anti–rabbit IgG (Jackson ImmunoResearch, 711-546-152), or CY 3-conjugated anti–mouse IgG (Jackson ImmunoResearch, 715-166-150), used for 1 hour at room temperature. Nuclei were stained with Nunc blue (Invitrogen, R37606). All staining images were taken with a Zeiss Axio observer Z1 with a 20× or 40× objective.
Swain S.M., Romac J.M., Vigna S.R, & Liddle R.A. (2022). Piezo1-mediated stellate cell activation causes pressure-induced pancreatic fibrosis in mice. JCI Insight, 7(8), e158288.
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2

Immunofluorescence Analysis of Piezo1 and Piezo2

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For the immunofluorescence analysis of Piezo1 and Piezo2, the cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 (Merck, #X100) for 10 min, blocked with 1% BSA (Sigma-Aldrich #A7030) for 30 min. Primary antibodies against Piezo1 (Alomone Labs, #APC-087) and Piezo2 (Alomone Labs, #APC-090) were incubated in PBS for 3 h at room temperature. Goat anti-rabbit secondary antibody, P conjugate, was used at a concentration of μg/mL in phosphate buffered saline containing 0.2% BSA for 1 h at room temperature. Nuclei were stained with DAPI. The neuronal component was stained with Tuji-1 (Sigma-Aldrich, #T3952)). The images were captured at 40X magnification using Eclipse Series TiE, equipped with a C1 confocal.
Falleroni F., Bocchero U., Mortal S., Li Y., Ye Z., Cojoc D, & Torre V. (2022). Mechanotransduction in hippocampal neurons operates under localized low picoNewton forces. iScience, 25(2), 103807.
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3

Immunofluorescence Staining of PIEZO1

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Cells seeded in flow chambers were fixed with 4% formaldehyde in PBS (Carl Roth 3105.2) for 20 min at room temperature and subsequently permeabilized with 0.1% Triton-X (Axonlab 10029070) and 0.5% bovine albumin serum (BSA, VWR P6154) in PBS for 10 min. Samples were incubated with specific primary antibodies in PBS with 3% BSA for 1 h and afterwards with a secondary fluorescently-labelled antibody. Between every step samples were washed three times with PBS. Primary antibodies were used against PIEZO1 (Novus Biologicals NBP1-78446 for human cells, 1:25; Alomone Labs APC-087 for rat cells, 1:300). Actin filaments were stained with Alexa Fluor 568 phalloidin (Thermo Fisher Scientific A12380, 1:200) and cell nuclei with NucBlue (Thermo Fisher Scientific R37605). Alexa Fluor 488 conjugated donkey anti-rabbit (Thermo Fisher Scientific A-21206, 1:100) was used as a secondary antibody. Immunofluorescence images were acquired with the iMic spinning disk confocal microscope using an oil-immersion 60x (N.A. 1.35) objective.
Passini F.S., Jaeger P.K., Saab A.S., Hanlon S., Chittim N.A., Arlt M.J., Ferrari K.D., Haenni D., Caprara S., Bollhalder M., Niederöst B., Horvath A.N., Götschi T., Ma S., Passini-Tall B., Fucentese S.F., Blache U., Silván U., Weber B., Silbernagel K.G, & Snedeker J.G. (2021). Shear-stress sensing by PIEZO1 regulates tendon stiffness in rodents and influences jumping performance in humans. Nature biomedical engineering, 5(12), 1457-1471.
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4

Immunofluorescence Staining of PIEZO1

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Cells seeded in flow chambers were fixed with 4% formaldehyde in PBS (Carl Roth 3105.2) for 20 min at room temperature and subsequently permeabilized with 0.1% Triton-X (Axonlab 10029070) and 0.5% bovine albumin serum (BSA, VWR P6154) in PBS for 10 min. Samples were incubated with specific primary antibodies in PBS with 3% BSA for 1 h and afterwards with a secondary fluorescently-labelled antibody. Between every step samples were washed three times with PBS. Primary antibodies were used against PIEZO1 (Novus Biologicals NBP1-78446 for human cells, 1:25; Alomone Labs APC-087 for rat cells, 1:300). Actin filaments were stained with Alexa Fluor 568 phalloidin (Thermo Fisher Scientific A12380, 1:200) and cell nuclei with NucBlue (Thermo Fisher Scientific R37605). Alexa Fluor 488 conjugated donkey anti-rabbit (Thermo Fisher Scientific A-21206, 1:100) was used as a secondary antibody. Immunofluorescence images were acquired with the iMic spinning disk confocal microscope using an oil-immersion 60x (N.A. 1.35) objective.
Passini F.S., Jaeger P.K., Saab A.S., Hanlon S., Chittim N.A., Arlt M.J., Ferrari K.D., Haenni D., Caprara S., Bollhalder M., Niederöst B., Horvath A.N., Götschi T., Ma S., Passini-Tall B., Fucentese S.F., Blache U., Silván U., Weber B., Silbernagel K.G, & Snedeker J.G. (2021). Shear-stress sensing by PIEZO1 regulates tendon stiffness in rodents and influences jumping performance in humans. Nature biomedical engineering, 5(12), 1457-1471.
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5

Immunohistochemical Detection of Piezo1 in Bladder

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For IHC, bladder sections stained with H&E were incubated overnight at 4 °C with primary antibody against Piezo1 (Alomone labs, APC-087, 1:100). Phosphate-buffered saline (PBS) was substituted for the primary antibody as a negative control. Following incubation with primary antibody, the sections were incubated with a streptavidin–biotin peroxidase (SP)-conjugated secondary goat anti-rabbit IgG from a standard SP kit (Zhongshan Co., SPN-9002) for 30 min at 37 °C. Finally, the sections were stained with diaminobenzidine (DAB) (Zhongshan Co., ZLI-9019) and counterstained with hematoxylin. All sections were visualized and photographed with an optical microscope (Olympus) and evaluated by two individuals in a double-blind manner.
Liu Q., Sun B., Zhao J., Wang Q., An F., Hu X., Yang Z., Xu J., Tan M, & Li L. (2018). Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis. Experimental & Molecular Medicine, 50(5), 60.
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