Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin-streptomycin solution, 0.25% trypsin/EDTA, and phosphate-buffered saline were purchased from Gibco. Fetal bovine serum (FBS) was purchased from Biological Industries. Rabbit monoclonal anti-Mfn1(Cat#: 14739S), rabbit monoclonal anti-Mfn2(Cat#: 9482S), rabbit monoclonal anti-Opa1(Cat#: 80471S), rabbit polyclonal anti-Drp1(Phospho Ser616 Cat#: 4494S), rabbit monoclonal anti-AMPKα (Phospho Thr172 Cat#: 2535S), and rabbit monoclonal anti-β-actin(Cat#: 4970T) were purchased from Cell Signaling Technology. Rabbit monoclonal anti-Fis1(Cat#: ab156865) and rabbit monoclonal anti-DRP1(Cat#: ab184247) were purchased from Abcam. DMSO, Mito-Tracker, BCA Protein Quantification Kit, RIPA lysis buffer, protease inhibitor cocktail, phosphatase inhibitors, SDS-PAGE Gel Preparation Kit, and other Western blotting reagents were purchased from Yeasen (Shanghai, China). Annexin V-Alexa Fluor 647/PI Apoptosis Detection Kit was purchased from Fcmacs (Nanjing, China). The mitochondrial membrane potential assay kit with JC-1 was purchased from Beyotime (Nanjing, China). Unless otherwise indicated, all other chemicals and reagents were purchased from Sigma-Aldrich.
Mito tracker
Manufactured by Yeasen
Sourced in China
The Mito-Tracker is a lab equipment designed to detect and analyze mitochondria within cells. It utilizes fluorescent dyes to specifically label and visualize mitochondria, allowing researchers to study their structure, distribution, and function within the cellular environment.
Automatically generated - may contain errors
Lab products found in correlation
Fluo 4 am, by Yeasen (2 mentions)
Protease inhibitor cocktail, by Yeasen (1 mentions)
Rabbit monoclonal anti β actin, by Cell Signaling Technology (1 mentions)
Phosphatase inhibitor, by Yeasen (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Rabbit monoclonal anti opa1, by Cell Signaling Technology (1 mentions)
Rabbit monoclonal anti mfn2, by Cell Signaling Technology (1 mentions)
Sds page gel preparation kit, by Yeasen (1 mentions)
Dimethyl sulfoxide (dmso), by Yeasen (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Ripa lysis buffer, by Yeasen (1 mentions)
Mitochondrial membrane potential assay kit with jc 1, by Beyotime (1 mentions)
Bca protein quantification kit, by Yeasen (1 mentions)
Annexin 5 alexa fluor 647 pi apoptosis detection kit, by Fcmacs (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Yeasen (1 mentions)
Annexin 5 fitc pi, by Yeasen (1 mentions)
Mapk family antibodysampler kit 9926t, by Cell Signaling Technology (1 mentions)
7 protocols using mito tracker
1
Mitochondrial Dynamics Regulation in Cell Apoptosis
2
Multicolor Fluorescence Microscopy for Cellular Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three additional fluorescence probes were used in this study, apart from Calcein-AM/PI and FDA, mentioned above. Images were obtained using a confocal microscope (Nikon). The cells were incubated for 30 min with 5 μM CellROX (50103ES50; Yeasen Biotechnology (Shanghai) Co. Ltd. Shanghai, China) to detect oxidative stress in them. CellROX gets localized in the cytoplasm and can react with reactive oxygen species (ROS) to turn bright fluorescent (ex/em, ∼ 545/565 nm). The cells were incubated for 30 min with 200 μM Mitotracker (40740ES50; Yeasen) or 15 μM JC-1(HY-15534; MCE) to detect oxidative stress. The cell-permeant Mitotracker® probes contain a mildly thiol-reactive chloromethyl moiety for mitochondrial labeling. Fluorescence intensity depends on the mitochondrial membrane potential (ex/em, ∼579/599 nm). JC-1 exhibits potential-dependent accumulation in the mitochondria, indicated by a fluorescence emission shift from green to red. While red fluorescence indicates a normal mitochondrial membrane potential, green fluorescence reflects an alteration of the latter (ex/em, ∼510/527 nm).
3
Multimodal Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT, calcein-AM, EthD-1, AnnexinV-FITC/PI, ER-tracker, Mito-Tracker, Fluo-4-AM,
DAPI, Hoechst 33342, H2DCF-DA, MEQ, and BCECF-AM were purchased from Yeasen
(Shanghai, China). Z-VAD-fmk, 3-methyladenine (3-MA), bafilomycinA1 (Baf),
cycloheximide (CHX), N-acetylcysteine (NAC), reduced
glutathione (GSH), SB203580, U0126, SP600125, ruthenium red (RR), and disodium
4,4′-diisothiocyanato-2,2′-stilbenedisulfonate hydrate (DIDS) were obtained from
Selleck Chemicals (Houston, TX). The following antibodies were used: Cleaved
Caspase Antibody Sampler Kit 9929T (cleaved caspase-3, -7, and -9 and cleaved
poly-ADP-ribose polymerase [PARP]), Procaspase Antibody Sampler Kit 12742T
(Caspase-3, -7, and -9; PARP), Bcl-2 (3498T), Bax (2772T), Beclin-1(3495T),
LC3I/II (12741T), p62(5114S), ER Stress Antibody Sampler Kit 9956T (calnexin,
BiP, IRE1α, CHOP), ubiquitin (43124S), XBP1s (83418S), MAPK Family Antibody
Sampler Kit 9926T (ERK 1/2, p38, JNK), Phospho-MAPK Family Antibody Sampler Kit
9910T (phospho-ERK1/2, phospho-p38, phospho-JNK, Rabbit IgG HRP, Mouse IgG HRP),
GAPDH (5174S), and chloride intracellular channel-1 (CLIC1; 53424S) were
purchased from Cell Signaling Technology (Danvers, MA). Alexa Fluor 647
AffiniPure Goat anti-Rabbit immunoglobulin G (IgG; H + L) (cat: FMS-RBaf64701)
was obtained from FCMRCS (Nanjing, China).
DAPI, Hoechst 33342, H2DCF-DA, MEQ, and BCECF-AM were purchased from Yeasen
(Shanghai, China). Z-VAD-fmk, 3-methyladenine (3-MA), bafilomycinA1 (Baf),
cycloheximide (CHX), N-acetylcysteine (NAC), reduced
glutathione (GSH), SB203580, U0126, SP600125, ruthenium red (RR), and disodium
4,4′-diisothiocyanato-2,2′-stilbenedisulfonate hydrate (DIDS) were obtained from
Selleck Chemicals (Houston, TX). The following antibodies were used: Cleaved
Caspase Antibody Sampler Kit 9929T (cleaved caspase-3, -7, and -9 and cleaved
poly-ADP-ribose polymerase [PARP]), Procaspase Antibody Sampler Kit 12742T
(Caspase-3, -7, and -9; PARP), Bcl-2 (3498T), Bax (2772T), Beclin-1(3495T),
LC3I/II (12741T), p62(5114S), ER Stress Antibody Sampler Kit 9956T (calnexin,
BiP, IRE1α, CHOP), ubiquitin (43124S), XBP1s (83418S), MAPK Family Antibody
Sampler Kit 9926T (ERK 1/2, p38, JNK), Phospho-MAPK Family Antibody Sampler Kit
9910T (phospho-ERK1/2, phospho-p38, phospho-JNK, Rabbit IgG HRP, Mouse IgG HRP),
GAPDH (5174S), and chloride intracellular channel-1 (CLIC1; 53424S) were
purchased from Cell Signaling Technology (Danvers, MA). Alexa Fluor 647
AffiniPure Goat anti-Rabbit immunoglobulin G (IgG; H + L) (cat: FMS-RBaf64701)
was obtained from FCMRCS (Nanjing, China).
4
Mitochondrial Imaging in Primary Microglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After OGD/R, primary microglial cells were stained with 200 nM mitotracker (YEASEN) at normal cell culture conditions for ≈30 min. After mitotracker staining, cells were immediately harvested on coverslip and images of fluorescence were captured with a confocal microscope (OLYMPUS, FV1200, Japan).
5
Evaluating Oocyte Mitochondrial Function
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the ROS content, mitochondrial membrane potential and mitochondrial distribution of oocytes, the procedure was conducted according to the protocols with the ROS kit (50101ES01, Yeasen, Shanghai, China), the JC-1 assay kit (40706ES60, Yeasen, Shanghai, China) and the MitoTracker (40740ES50, Yeasen, Shanghai, China), respectively. First, MII oocytes were collected from ampulla of fallopian tube and immediately washed with M2 medium 3 times. Then oocytes were transferred into M2 medium containing staining solution and incubated at 37 C in the dark for 30 min. After that, oocytes were washed with M2 medium to remove the excess staining solution. All of the live oocytes were observed by a laser scanning confocal microscope (LSM800, Zeiss, Germany). The relative fluorescence intensity was calculated by software of ZEN.
6
Visualizing NEU1-ALK5 Interaction in HK-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HK-2 cells were plated on glass bottom dish. Transfections were carried out using the lip3000 reagent, with pBiFC-NEU1-VC155 and pBiFC-ALK5-VN173 plasmid. Cells were stimulated with or without TGFβ (10 ng/ml, PeproTech) for 24 h after transfection 24 h, and were stained with Hoechst 33342 (C1028, Beyotime) to label nuclear DNA or with Lyso-Tracker (C1046, Beyotime), Mito-Tracker (40740ES50, Yeasen), and CM-Tracker (C1036, Beyotime) to label lysosome, mitochondrion, and cell plasma membrane, respectively. For the control group, cells were transfected with pBiFC-HA-NEU1-VC155 and pBiFC-VN173 plasmids or pBiFC-ALK5-VN173 and pBiFC-VC155 plasmids. Fluorescence signal amplification was observed using the Zeiss LSM800 confocal laser scanning microscopy.
7
Visualizing Calcium in Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect calcium in cytoplasm and mitochondria, macrophages in glass bottom dishes were incubated with 1 μM Fluo4 AM and 200 nM Mitotracker (Yeasen Biotechnology) at 37°C for 30 minutes in the dark, respectively, and then observed with confocal microscopy.
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!