Eos kiss x6i

Manufactured by Canon

Sourced in Japan

The EOS Kiss X6i is a digital single-lens reflex (DSLR) camera. It features a 18.0-megapixel APS-C CMOS sensor and DIGIC 4 image processor. The camera supports Full HD 1080p video recording at 30 fps. It has a 3.0-inch LCD screen and offers various shooting modes and connectivity options.

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9 protocols using eos kiss x6i

Immunoblotting for TUBB3 Expression

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Anti-E-cadherin, anti-ATP-binding cassette subfamily B, member 1 (ABCB1), anti-class III beta-tubulin (TUBB3) and anti-beta-actin antibodies were purchased from Cell Signaling Technology (Beverly, MA).
Preparation of total cell lysates and immunoblotting was performed as previously described [17 (link)]. Cells were cultured without erlotinib until subconfluency, and media was changed to RPMI with 10% FBS containing DMSO or 1 μM entinostat. After 72 hours, cells were rinsed with phosphate-buffered saline (PBS), lysed in sodium dodecyl sulfate (SDS) buffer and homogenized. Approximately 30 μg of total cell lysate protein was subjected to SDS polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA). After blocking with 2.5% nonfat dry milk and 2.5% bovine serum albumin in PBS, membranes were incubated with primary antibodies (1:1000) overnight, washed with PBS, reacted with secondary antibody (1:1000), treated with ECL solution (GE Healthcare, Fairfield, CT). Chemiluminescence was detected by EOS Kiss X6i (Canon, Tokyo, Japan). Expression values of TUBB3 relative to beta-actin were determined using Just TLC software (Sweday, Lund, Sweden).

Mizuuchi H., Suda K., Sato K., Tomida S., Fujita Y., Kobayashi Y., Maehara Y., Sekido Y., Nishio K, & Mitsudomi T. (2015). Collateral Chemoresistance to Anti-Microtubule Agents in a Lung Cancer Cell Line with Acquired Resistance to Erlotinib. PLoS ONE, 10(4), e0123901.

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Quantitative Protein Assay on μPAD

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First, 15 μL of a 250 mM citrate buffer solution (pH 1.8) was introduced into the buffer zone and was evaporated using a dryer for 2 min. Then, 15 μL of a 9 mM TBPB solution in 95% ethanol was introduced into the buffer zone and was evaporated using a dryer for 5 min. Finally, 3.5 μL of BSA solutions of different concentrations was separately spotted onto the eight detection zones. Colorimetry was used as the detection method. The distance between the μPAD and a digital camera (EOS Kiss X6i, Canon, Tokyo, Japan) was ca. 20 cm, and images of the PADs were taken using a digital camera under fluorescent lighting conditions. The color information was measured with an image analysis software program (ImageJ Ver. 1.48).

Komatsu T., Maeki M., Ishida A., Tani H, & Tokeshi M. (2018). Characteristics of Microfluidic Paper-based Analytical Devices Fabricated by Four Different Methods. Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 34(1).

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Profiling RTK Phosphorylation Levels

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A human Phospho‐RTK Array Kit (R&D Systems, Minneapolis, MN, USA) was used to detect the relative level of tyrosine phosphorylation of 49 receptor tyrosine kinases as described previously.19 Briefly, cells were lysed by NP‐40 lysis buffer according to the manufacturer's protocol. Membranes with antibodies spotted against 49 phospho‐RTK were treated with blocking buffer and incubated with 450 μg of cell lysate overnight at 4°C. The arrays were then washed, incubated with an HRP‐conjugated phospho‐tyrosine detection antibody, treated with ECL solution, and detected with EOS Kiss X6i (Canon, Tokyo, Japan).

Mizuuchi H., Suda K., Murakami I., Sakai K., Sato K., Kobayashi Y., Shimoji M., Chiba M., Sesumi Y., Tomizawa K., Takemoto T., Sekido Y., Nishio K, & Mitsudomi T. (2016). Oncogene swap as a novel mechanism of acquired resistance to epidermal growth factor receptor‐tyrosine kinase inhibitor in lung cancer. Cancer Science, 107(4), 461-468.

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Western Blot Analysis of Signaling Proteins

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Cells were cultured in RPMI 1640 with 10% FBS until subconfluency and media was changed to new media with indicated concentration of test drugs. After 24 h, cells were rinsed with PBS, lysed in SDS buffer, and homogenized using a scraper. Approximately 20 μg to total cell lysates were subjected to SDS polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Bio‐Rad, Hercules, CA, USA). After blocking with Western BLoT Blocking Buffer Protein Free (TaKaRa) or PBS containing 2.5% skimmed milk and 2.5% BSA, membranes were incubated with primary antibodies (1:2000) overnight, washed with PBS containing tween‐20 (PBS‐T), reacted with secondary antibody (1:5000), and washed with PBS‐T again and treated with Western BLoT Quant HRP Substrate (TaKaRa). Chemiluminescence was detected with by EOS Kiss X6i (Canon). Anti‐phospho‐EGFR, anti‐EGFR, anti‐phospho‐MET, anti‐MET, anti‐phospho‐AKT, ant‐AKT, anti‐phospho‐ERK, anti‐ERK, anti‐PARP, anti‐cleaved PARP, anti‐GAPDH and anti‐beta‐actin antibodies were all purchased from Cell Signaling Technologies (Danvers, MA, USA).

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Immunostaining of hiPS-Derived Cardiomyocytes

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For the characterization of hiPS-CMs by immunostaining, samples were washed with PBS, fixed with 4% paraformaldehyde (PFA) (Muto Pure Chemicals Co., Ltd.), permeabilized with 0.1% Triton X-100 for 20 min, and blocked with 2.5% bovine serum albumin (BSA) (Sigma-Aldrich) overnight. Afterward, we incubated the samples with primary antibodies at 4 °C overnight and then with secondary antibodies at room temperature for 2 h. For actin staining, we used 0.1% Alexa Fluor 647-conjugated phalloidin (Thermo Fisher Scientific, Inc.) as the primary antibody, and did not use secondary antibodies. For α-actinin staining, we used 0.1% monoclonal anti-α-actinin antibody (Sigma-Aldrich) as the primary antibody, and 0.1% goat anti-mouse IgG antibody (Thermo Fisher Scientific, Inc.) as the secondary antibody. Following this, we rinsed them with PBS and stained cell nuclei with 0.1% Hoechst 33342 (Invitrogen).
In order to observe the samples, we used a digital camera with a macro lens (EOS Kiss X6i; Canon) for bright-field images, a microscope (IX71N; Olympus) for bright-field and fluorescence microscopy, and a laser microscope (LSM780; Zeiss) for confocal microscopy.

Morimoto Y., Mori S., Sakai F, & Takeuchi S. (2016). Human induced pluripotent stem cell-derived fiber-shaped cardiac tissue on a chip. Lab on a chip, 16(12).

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Morphological Development of Jellyfish

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Observations of early development from fertilized eggs to planulae using a stereomicroscope were conducted at 2, 4, 6, and 24 h after the collection of fertilized eggs. Morphological observations of polyps, strobilae, ephyrae, metaephyrae, and medusae were similarly conducted, and when animals relaxed under the microscope, images were taken from directly above using a digital camera (EOS Kiss X6i, Canon, Tokyo, Japan) mounted on the stereomicroscope or biological microscope. The mouth disc diameter of the polyps and strobilae and the central disc diameters of the ephyrae [20 (link)] and medusae were measured from photo images using Image J software [21 (link)].

Miyake H., Honda S., Nishikawa J, & Yusoff F.M. (2021). Life Cycle of Edible Jellyfish Acromitus hardenbergi Stiasny, 1934 (Scyphozoa: Rhizostomeae) Inhabiting a Brackish-Water Environment. Animals : an Open Access Journal from MDPI, 11(7), 2138.

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Embryonic Development Monitoring Protocol

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Shortly after spawning, eggs were collected and left to stand for a few minutes to distinguish between fertilised (living) eggs and unfertilised (dead) eggs; fertilised eggs float, whereas unfertilised eggs sink to the bottom. The floating eggs on the water surface were carefully withdrawn using a plastic pipette; groups of ~50 eggs each were transferred to a plastic Petri dish (15-cm diameter), filled with sterile seawater passed through a 0.22-µm filter and incubated at 20 °C for up to 4 d. To maintain water quality, dead embryos were removed and the seawater was replaced after 3 and 24 h incubation. The embryos were observed under stereomicroscope (Leica M60; Leica Microsystems); their images were captured using digital camera (EOS Kiss X6i; Canon Inc.).

Sakaguchi K., Yoneda M., Sakai N., Nakashima K., Kitano H, & Matsuyama M. (2019). Comprehensive Experimental System for a Promising Model Organism Candidate for Marine Teleosts. Scientific Reports, 9, 4948.

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Biohybrid Robot Imaging Techniques

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To observe the flexible substrate with anchors and the skeletal muscle tissue on the substrate, we used a digital camera with a macro lens (EOS Kiss X6i; Canon Inc.,) to obtain a bright-field image and a confocal microscope (LSM780; Carl Zeiss). For observing the biohybrid robot, we used a macroscopic scope (MVX10; Olympus Corp.) to obtain fluorescent images of the robot and bright-field images of the collagen structure containing the skeletal muscle tissue. To obtain bright-field images and fluorescent images of the skeletal muscle tissue, we used a microscope (IX71N, Olympus Corp.).

Morimoto Y., Onoe H, & Takeuchi S. (2020). Biohybrid robot with skeletal muscle tissue covered with a collagen structure for moving in air. APL Bioengineering, 4(2), 026101.

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Monitoring Powdery Mildew Spread in Melon Seedlings

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Mature KMP-6N conidia were inoculated onto the well-developed young leaves of 14-day-old melon seedlings using the methods described above [23] (link). In each replicate, a KMP-6N-inoculated seedling was placed at the centre of a multi-pack tray, surrounded by 48 healthy 7-day-old melon seedlings, and maintained in a temperature-controlled room (25 ± 1 • C; 45-55% RH), with continuous illumination of 60 µmoL m -2 s -1 under red or blue LEDs or 22.2 µmoL m -2 s -1 under white fluorescent lamps. A continuous air current of 1.0 m s -1 [39] (link) was provided to the seedlings using an electric fan positioned at a 1 m distance from the multi-pack tray. The current study photographed the 48 healthy seedlings at 24 h intervals using an EOS KISSX6i digital camera (Canon, Tokyo, Japan) and observed for 21 days to analyse the spread of powdery mildew to healthy melon seedlings. Finally, the fungal colonies appeared on leaves of cotyledon-stage seedlings at 21 days after treatments were checked with photographs and eyes.

Suzuki T., Iwasaki S., Hisazumi H., Miyamoto A., Ogami H., Takikawa Y., Kakutani K., Matsuda Y., & Nonomura T. (2022). Inhibitory Effects of Blue Light-Emitting Diode Irradiation on Podosphaera xanthii Conidial Release and Infection of Melon Seedlings. Agriculture, 12(2), 198-198.

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