Magcapture exosome isolation kit ps

Manufactured by Fujifilm

Sourced in Japan

The MagCapture Exosome Isolation Kit PS is a laboratory tool designed for the isolation and purification of exosomes from biological samples. It utilizes magnetic beads to capture and separate exosomes from the sample, enabling their extraction for further analysis or research purposes.

Automatically generated - may contain errors

Lab products found in correlation

Exoquick solution, by System Biosciences (3 mentions) Exo fbs 50a 1, by System Biosciences (2 mentions) Exoquick exosome precipitation solution, by System Biosciences (2 mentions) Exosome depleted fbs, by System Biosciences (2 mentions) Collagen type 1, by Corning (1 mentions) Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions) Insulin transferrin selenium solution, by Thermo Fisher Scientific (1 mentions) Recombinant rat ifnγ, by R&D Systems (1 mentions) Dmem f12, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Nichirei Biosciences (1 mentions) Vp sfm, by Thermo Fisher Scientific (1 mentions) Amicon ultra 100k, by Merck Group (1 mentions) Gw4869, by Cayman Chemical (1 mentions) Millex gv filter, by Merck Group (1 mentions) Nanosight lm10, by Malvern Panalytical (1 mentions) 0.22 μm spin filter, by Agilent Technologies (1 mentions) Universal sybr select master mix, by Thermo Fisher Scientific (1 mentions) Revertra ace qpcr master mix, by Toyobo (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Lightcycler 96, by Roche (1 mentions)

25 protocols using magcapture exosome isolation kit ps

Isolation and Analysis of Rat CP Cell Exosomes

Check if the same lab product or an alternative is used in the 5 most similar protocols

The rat CP cell line (The European Collection of Authenticated Cell Cultures, London, UK) were used for culturing. CP cells were seeded at a density of 0.8 × 10⁴ cells/cm² in a 24-well plate coated with type 1 collagen (Corning, Corning, NY, USA). Cells were cultured in DMEM/F-12 (Sigma-Aldrich, Saint Louis, MO, USA), 10% FBS (CCB, NICHIREI BIOSCIENCE, Tokyo, Japan), 10 mg/ml recombinant rat epidermal growth factor protein (R&D Systems), 1% Insulin/Transferrin/Selenium Solution (Thermo Fisher Scientific), and 1% penicillin streptomycin (PS) (Thermo Fisher Scientific) (Battle et al., 2000 (link)). Cells were maintained in a 5% CO₂ incubator at 37 °C. At 3 days after seeding when each cultured cell type had reached 90–100% confluence, the conventional medium was replaced with 10% EXO-FBS-50A-1 (System Biosciences, Palo Alto, CA, USA) medium with several concentrations of recombinant rat IFN-γ (0 or 0.5 ng/ml; R&D Systems). At 24 h after incubation, the culture media were collected and the exosomes were isolated using the MagCapture Exosome Isolation Kit PS (FUJIFILM Wako Pure Chemical Corp, Osaka, Japan).
miRNA isolation, cDNA synthesis, and RT-PCR were performed as described above. The relative expression of miR-146a was calculated using 2^−ΔΔCt with cel-miR-39 as an external spiked control.

Nakano M., Kubota K., Hashizume S., Kobayashi E., Chikenji T.S., Saito Y, & Fujimiya M. (2020). An enriched environment prevents cognitive impairment in an Alzheimer’s disease model by enhancing the secretion of exosomal microRNA-146a from the choroid plexus. Brain, Behavior, & Immunity - Health, 9, 100149.

+ Open protocol

+ Expand

Isolation of Extracellular Vesicles using TIM-4 Affinity

Check if the same lab product or an alternative is used in the 5 most similar protocols

Prior to EV isolation, cells were sub-cultured to 20, 10 cm dishes for a total culture volume of 200 ml. Cells were incubated in serum-free media for 24 h before isolation. TIM-4, a phospholipid phosphatidylserine present on the surface of the extracellular vesicle, can be used to isolate EVs from the serum-free media. To isolate EVs by the TIM-4 affinity method, a MagCapture Exosome Isolation Kit PS (Wako, Japan) was used according to the manufacturer’s instructions. In brief, 0.6 mg of streptavidin magnetic beads, bound with 1 μg of biotinylated mouse Tim4-Fc, was added to 10K filtered supernatant and the mixture was rotated overnight at 4 °C. The beads were washed three times with 1 ml of washing buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.0005% Tween20, 2 mM CaCl₂), and the bound EVs were eluted with elution buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA).

Glady A., Vandebroek A, & Yasui M. (2021). Human keratinocyte-derived extracellular vesicles activate the MAPKinase pathway and promote cell migration and proliferation in vitro. Inflammation and Regeneration, 41, 4.

+ Open protocol

+ Expand

Prostate Cancer Exosome Isolation Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

After filtration, 80 mL of cell supernatant was filtered using an Amicon^® Ultra filter device by centrifugation at 4000 g for 30–80 min, and PBS was added to the filter to continue centrifugation to obtain a suspension of 1.5 mL of prostate cancer exosomes (PCa-exos) [19 (link)]. A pipette was inserted into the bottom of the filter, and the sample was swept back and forth at the bottom to ensure complete exosome recovery. Afterwards, these exosomes were further purified using the Magcapture™ Exosome Isolation Kit PS (Fujifilm Wako, Osaka, Japan) according to the instructions. The obtained exosomes were stored at − 20 °C.

Peng Y., Zhao M., Hu Y., Guo H., Zhang Y., Huang Y., Zhao L., Chai Y, & Wang Z. (2022). Blockade of exosome generation by GW4869 inhibits the education of M2 macrophages in prostate cancer. BMC Immunology, 23, 37.

+ Open protocol

+ Expand

EV Isolation from Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

The BHK/JM-PnL and BHK/DN-PnL cells in the culture plates were rinsed with PBS and then cultured in 10% VP-SFM (Thermo Fisher Scientific)–90% MEM supplemented with NEAA (VPSFM-MEM). Then, the culture supernatants were collected daily and subjected to centrifugation at 3000 rpm for 5 min to remove cell debris. The cleared culture fluids were concentrated by about 100-fold with the Amicon Ultra 100 K (Merck, Burlington, MA, USA). Approximately 1 mL of the concentrated culture fluids were subjected to EV isolation by using the MagCapture™ exosome isolation kit PS (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) according to the manufacturer’s instruction except that the addition of the beads was repeated three times for each culture fluid preparation. All of the EVs eluted from the three batches of beads were combined and used as the EV sample derived from each fluid preparation. After EV isolation with the beads, the remaining concentrated culture fluids were used as EV-depleted samples in some analyses.
In some experiments where EV production needs to be inhibited, GW4869 (7.5 µM in DMSO; Cayman Chemical, Ann Arbor, MI, USA) was added 24 h before changing the media to VPSFM-MEM, which also contained GW4869.

Ishikawa T., Narita K., Matsuyama K, & Masuda M. (2024). Dissemination of the Flavivirus Subgenomic Replicon Genome and Viral Proteins by Extracellular Vesicles. Viruses, 16(4), 524.

+ Open protocol

+ Expand

Purification of Urine Extracellular Vesicles

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purification of EVs from urine with Tim4-affinity beads (Nakai et al., 2016 (link)) was performed using the MagCapture Exosome Isolation Kit PS (Cat# 293-77601 FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) following the manufacturer’s protocol. Urine samples were centrifuged at 1,200 × g for 20minat 4°C to remove cell debris and urine salts and then centrifuged at 10,000 × g for 30 min at 4°C to remove the large EVs such as apoptotic bodies. Streptavidin magnetic beads, bound with biotinylated Tim4, were added to 1 mL of the supernatant supplemented with 2 mM CaCl₂, and the mixture was rotated for 1hat 20–25°C. The beads were washed three times with 1 mL washing buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.0005% Tween 20, 2 mM CaCl₂), and the bound uEVs were eluted with an elution buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA).

Takizawa K., Ueda K., Sekiguchi M., Nakano E., Nishimura T., Kajiho Y., Kanda S., Miura K., Hattori M., Hashimoto J., Hamasaki Y., Hisano M., Omori T., Okamoto T., Kitayama H., Fujita N., Kuramochi H., Ichiki T., Oka A, & Harita Y. (2022). Urinary extracellular vesicles signature for diagnosis of kidney disease. iScience, 25(11), 105416.

+ Open protocol

+ Expand

Isolation and Characterization of sEVs from HEK293T Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

sEVs of the HEK293T cell were isolated according to the protocol that was reported previously (Lim et al., 2020 (link), 2021 (link)). After 4 days of culturing, cell‐conditioned medium was collected and subjected to differential centrifugation to remove cells, debris, and large EVs. Then, the supernatant was filtered through a 0.22‐μm Millex‐GV filter (Merck Millipore, MA, USA) to generate a final pre‐cleared cell conditioned medium, known as the 10K sup. After that, a MagCapture Exosome Isolation Kit PS (Wako, Osaka, Japan) was used following to the manufacturer's manual to purify the sEVs from the 10K sup. The purified sEVs were kept in a dialysis membrane (3500 MWCO) for overnight dialysis in PBS. NanoSIGHT LM10 (Malvern Panalytical, Malvern, UK) was used to perform nanoparticle tracking analysis (NTA) to measure the sEV concentration. The sEV purification process follows the Minimal Information for Studies of Extracellular Vesicle 2018 (MISEV2018) guidelines (Théry et al., 2018 (link)) because we uploaded the protocol to the EV‐TRACK knowledgebase in our earlier study (Lim et al., 2021 (link)) (EV‐TRACK ID: EV210256).

Sajidah E.S., Lim K., Yamano T., Nishide G., Qiu Y., Yoshida T., Wang H., Kobayashi A., Hazawa M., Dewi F.R., Hanayama R., Ando T, & Wong R.W. (2022). Spatiotemporal tracking of small extracellular vesicle nanotopology in response to physicochemical stresses revealed by HS‐AFM. Journal of Extracellular Vesicles, 11(11), 12275.

+ Open protocol

+ Expand

Purification of Rotavirus Vesicles

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 6-days-old BALB/c mouse pups were orally inoculated with 100-fold diluted rotavirus stock, stool was harvested twice a day from the diarrhea symptom onset to 5 dpi. Mouse pups’ bellies were gently pressed for stool collection at anuses, until no stool was released. The intestine content was also collected after pups were euthanized and intestines were surgically extracted. All rotavirus positive stool and the intestine content were suspended with 1× PBS buffer to obtain a 10%-20% (w/v) fecal suspension before being proceeded to a series of sequential centrifugations for preparing a clarified fecal solution. The sequential centrifugations were conducted for 10 min each with the speed of 500, 1,000, 2,000, 4,000, and 5,000 ×g at 10 °C. The fecal supernatant was transferred to a new tube after the centrifugation at 1,000, 4,000, and 5,000 ×g, and multiple rounds of centrifugation at 5,000 ×g were conducted when needed. The clarified fecal solution was then proceeded to vesicle purification using the MagCapture Exosome Isolation Kit PS (FUJIFILM Wako Pure Chemical, 293-77601), which uses TIM4-coated magnetic beads to selectively bind viral vesicles but not free viruses.^{1 (link)} Rotavirus vesicles were stored in the elution buffer of the kit at 4 °C for further use.

Zhang M., Ghosh S., Li M., Altan-Bonnet N, & Shuai D. (2022). Vesicle-Cloaked Rotavirus Clusters are Environmentally Persistent and Resistant to Free Chlorine Disinfection. Environmental science & technology, 56(12), 8475-8484.

+ Open protocol

+ Expand

Exosome Isolation from Serum Using MagCapture

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Exosome isolation from serum sample was performed using MagCapture Exosome Isolation Kit PS (Wako, 293-77601), according to the manufacturer’s instructions^{80 (link)}. Magnetic beads coated with Tim4 protein (which binds to Phosphatidylserine on the membrane surface of extracellular vesicles) enable purification of highly purified EVs.

Minami Y., Hoshino A., Higuchi Y., Hamaguchi M., Kaneko Y., Kirita Y., Taminishi S., Nishiji T., Taruno A., Fukui M., Arany Z, & Matoba S. (2023). Liver lipophagy ameliorates nonalcoholic steatohepatitis through extracellular lipid secretion. Nature Communications, 14, 4084.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation from CSF

Check if the same lab product or an alternative is used in the 5 most similar protocols

CSF samples were centrifuged at 1200× g for 20 min at 4 °C, and then the supernatant was centrifuged at 10,000× g for 30 min at 4 °C. The subsequent supernatant was then filtered in a 0.22 μm Spin-X centrifuge tube (#CLS8160 Corning, Corning, NY, USA), and then the EV fraction was separated from the flow-through using the MagCapture Exosome Isolation Kit PS (#293-77601 Fujifilm WAKO Pure Chemical Corporation, Tokyo, Japan), according to the manufacturer’s instructions. The separated EVs were filtered in a 0.45 μm Spin-X centrifuge tube (#CLS8162 Corning) to completely remove magnetic beads.

Muraoka S., Jedrychowski M.P., Yanamandra K., Ikezu S., Gygi S.P, & Ikezu T. (2020). Proteomic Profiling of Extracellular Vesicles Derived from Cerebrospinal Fluid of Alzheimer’s Disease Patients: A Pilot Study. Cells, 9(9), 1959.

+ Open protocol

+ Expand

Exosome-mediated GABA transfer in Caco-2 and SH-SY5Y cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Firstly, Caco-2 cells (1.4 × 10⁵ cells/mL) were cultured in DMEM containing 10% Exosome-depleted FBS media supplement heat inactivated (System Bioscience, Mountain View, CA, USA) and 500 or 1000 μM GABA. After 24 h of culture, the MagCapture Exosome Isolation Kit PS (FUJIFILM Wako Pure Chemical Corp.) was used to isolate exosomes from the media of Caco-2 cells, according to the manufacturer’s instructions. Exosomes were isolated from mouse serum using ExoQuick Exosome Precipitation Solution (System Biosciences, Palo Alto, CA, USA), according to the manufacturer’s instructions. SH-SY5Y cells (2.0 × 10⁵ cell/mL) were cultured for 24 h, and treated with exosomes (equivalent to 90 ng protein) derived from GABA-treated Caco-2 cells for 24 h.

Inotsuka R., Udono M., Yamatsu A., Kim M, & Katakura Y. (2021). Exosome-Mediated Activation of Neuronal Cells Triggered by γ-Aminobutyric Acid (GABA). Nutrients, 13(8), 2544.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!