The cells were fixed with 2% paraformaldehyde (PFA)/PBS (Wako) and methanol (Wako), blocked with Blocking One (Nacalai) for 45 min, and subsequently incubated with primary antibodies diluted in 5% Blocking One/PBST (Wako) at 4 °C overnight. The cells were washed in PBS and incubated with secondary antibodies diluted in 5% Blocking One/PBST for 1 h at room temperature (24–26 °C). Then, 4′,6-diamidino-2-phenylindole (DAPI; Sigma) was used to counterstain the nuclei. The samples were visualized and photographed with a BZ-710X (Keyence; Osaka, Japan) or Opera Phenix System (PerkinElmer; Waltham, MA, USA). The primary antibodies used for this study were: mouse anti-myosin heavy-chain (pan-MHC) monoclonal (MF20; 1:500; R&D; Minneapolis, MN, USA), mouse anti-dystrophin (Rod domain) monoclonal (DYS1; 1:20; Leica; Buffalo Grove, IL, USA), rabbit anti-STIM1 monoclonal (D88E10; 1:800; Cell Signaling Technology; Danvers, MA, USA), and mouse anti-Orai1 monoclonal (G-2; 1:5; Santa Cruz; Dallas, TX, USA) antibodies. Alexa Fluor 488-conjugated anti-mouse/rabbit and Alexa Fluor 647-conjugated anti-mouse/rabbit antibodies (1:500, Invitrogen) were used as secondary antibodies.
Bz 710x
Manufactured by Keyence
Sourced in Japan
The BZ-710X is a high-performance digital microscope designed for laboratory applications. It provides clear, high-resolution imaging capabilities for a variety of samples.
Automatically generated - may contain errors
Lab products found in correlation
Methanol, by Fujifilm (2 mentions)
Blocking one, by Fujifilm (2 mentions)
Alexa fluor 488 conjugated anti mouse rabbit, by Thermo Fisher Scientific (2 mentions)
Opera phenix system, by PerkinElmer (2 mentions)
Blocking one, by Nacalai Tesque (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Matrigel, by Corning (1 mentions)
My 32, by Merck Group (1 mentions)
5 protocols using bz 710x
1
Multicolor Immunofluorescence Imaging of Skeletal Muscle Proteins
2
Matrigel-based Tube Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Matrigel-based tube formation assay was performed as previously described. LECs were transfected with Dusp6 siRNA or non-targeting siRNA. Each well of a 24-well plate was coated with 100 μL growth factor-reduced Matrigel (Corning, Corning, NY, USA) with or without 1.6 μg CCN2, which was allowed to polymerize for 30 min at 37 °C. Then, LECs were seeded onto the coated wells at a density of 5 × 104 cells per well and cultured in 500 μL FBS-free and supplement-free EGM2 medium. After incubation for 8 h at 37 °C under 5% CO2, tube formation images were captured at 100 × magnification by microscopy (BZ-710X; Keyence) in three random fields. For quantification, the tube length and branching points were measured.
3
Cofilin-mRFP expression in rat neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
E18 rat neurons were infected on DIV 3 with adenoviruses (100 multiplicity of infection (moi)) for expression of cofilin-mRFP (WT) [29 (link)]. Adenoviruses were made using the AdEasy system, with viruses tittered using the E2a immunoassay [69 (link),70 (link),71 (link)]. At 6 DIV, rods were induced by a change of medium to that containing Aβd/t. Rods that formed after overnight incubation were visualized by live-cell imaging on an incubation stage (Tokai Hit, Shizuoka, Japan) of a fluorescence microscope (BZ-710X, Keyence Corp. of America, Itasca, IL, USA) and were followed at 4 min intervals for ~150 min in the presence and absence of the CCR antagonists, AMD3100 [50 nM] and RAP-103 [0.1 nM].
4
Immunostaining of Intracellular Organelles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 12 h after transfection, CAD cells were treated with 2 μg/ml brefeldin A (BFA) for 4 h. After washing with medium containing 100 μg/ml cycloheximide, cells were further incubated for the indicated time and immunostained with anti-GM130 antibody. Primary cultured mouse cortical neurons (in vitro, day 5) were transfected with plasmids by the calcium phosphate method and cultured for 8 h. Cells were fixed and immunostained with indicated antibodies. Fluorescence images were acquired with a fluorescence microscope (BZ-710X; Keyence, Osaka, Japan), and quantitative analysis and colocalization efficiency were analyzed with Fiji/ImageJ and the Colco2 Fiji plug-in (ImageJ-Fiji-ImgLib; http://Fiji.sc or http://imageJ.net ).
5
Immunofluorescence Staining of Myosin Subtypes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with 2% paraformaldehyde (PFA)/PBS (Wako), methanol (Wako), blocked with Blocking One (Nacalai) for 45 min, and subsequently incubated with primary antibodies diluted in 5% Blocking One/ PBST (Wako) at 4°C overnight. Cells were washed three times in PBS and incubated with secondary antibodies diluted in 5% Blocking One/PBST for 1 h at room temperature. DAPI (Sigma) was used to counterstain the nuclei. Samples were visualized and photographed with BZ-710X (Keyence, Osaka, Japan) or the Opera Phenix System (PerkinElmer, Waltham, MA). The antibodies used for this study were as follows: mouse anti-myosin heavy chain (pan-MHC) monoclonal (MF20; 1:500, R&D, Minneapolis, MN), mouse anti-skeletal myosin (FAST/MYH1&2), monoclonal (MY-32; 1:100, Sigma), mouse anti-Myosin-2 (MYH2) monoclonal (MABT840; 1:100, Millipore, Burlington, MA), rabbit anti-MYH3 polyclonal (1:100, Atlas antibodies, Bromma, Sweden), mouse anti-MYH7 monoclonal (A4.840; 1:200, Santa Cruz, Dallas, TX), rabbit anti-MYH8 polyclonal (1:100, Novus Biologicals, Littleton, CO), rabbit anti-sarcomeric α actinin polyclonal (1:500, Abcam, Cambridge, UK), mouse anti-dystrophin (Rod domain) monoclonal (DYS1; 1:20, Leica, Buffalo Grove, IL), Alexa Fluor 488-conjugated anti-mouse/rabbit, and Alexa Fluor 647-conjugated anti-mouse/rabbit antibodies (1:500, Invitrogen).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!