Anti rabbit igg dylight 488

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-rabbit IgG-DyLight 488 is a secondary antibody conjugated with the DyLight 488 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassays and imaging applications.

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Lab products found in correlation

Anti mouse igg dylight 594, by Thermo Fisher Scientific (3 mentions) Anti mouse igg dylight 800, by Thermo Fisher Scientific (2 mentions) Dapi fluoromount g, by Electron Microscopy Sciences (1 mentions) Fluoview 1000 confocal microscope, by Olympus (1 mentions) Mouse anti human ki 67, by Cell Signaling Technology (1 mentions) Cytoflex flow cytometry, by Beckman Coulter (1 mentions) E cadherin, by BD (1 mentions) Antibodies against caveolin 1, by BD (1 mentions) Recombinant human tgf β1, by R&D Systems (1 mentions) Sb431542, by Merck Group (1 mentions) Protein a g plus agarose, by Santa Cruz Biotechnology (1 mentions) N cadherin, by BD (1 mentions) Mg132, by Merck Group (1 mentions) Trastuzumab, by Roche (1 mentions) Hct116, by American Type Culture Collection (1 mentions) Sw480, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Chloroquine diphosphate salt, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Corning (1 mentions) Odyssey protein imaging system, by LI COR (1 mentions)

Close spelling variants of this product

DyLight 488 (158 citations) Anti‐rabbit Dylight 488 (4 citations) IgG Rabbit (4 citations)

7 protocols using anti rabbit igg dylight 488

Immunofluorescent Labeling of TRPM4 and Aquaporin-1

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Anterior chambers were fixed in 4% paraformaldehyde for 1 hour, cryoprotected in 15 and 30% sucrose gradients, embedded in Tissue-Tek^® O.C.T. (Sakura, 4583), and cryosectioned at 12 µm, as described (13 (link), 30 (link)). Sections were probed with a polyclonal rabbit TRPM4 antibody (1:100 (32 (link)); and aquaporin-1 mouse monoclonal antibody (1:1000; Santa Cruz Biotechnology Sc-25287). Secondary antibodies were anti-rabbit IgG DyLight 488 (Invitrogen, 35552) and anti-mouse IgG DyLight 594 (Invitrogen, 35511). Sections were coverslipped with DAPI-Fluoromount-G (Electron Microscopy Sciences, Hatfield, PA, 17984-24) and imaged with a confocal microscope. Images were acquired using identical (HV, gain, offset) parameters.

Yarishkin O., Phuong T.T., Vazquez-Chona F., Bertrand J., van Battenburg-Sherwood J., Redmon S.N., Rudzitis C.N., Lakk M., Baumann J.M., Freichel M., Hwang E.M., Overby D, & Križaj D. (2022). Emergent Temporal Signaling in Human Trabecular Meshwork Cells: Role of TRPV4-TRPM4 Interactions. Frontiers in Immunology, 13, 805076.

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Immunohistochemical Analysis of Piezo1 in Mouse Eyes

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C57BL/6J and Piezo1^P1-tdT (Piezo1^tm1.1Apat) mice were killed by isoflurane inhalation followed by cervical dislocation, after which eyes were enucleated. Anterior chambers were fixed in 4% para-formaldehyde for one hour, cryoprotected in 15 and 30% sucrose gradients, embedded in Tissue-Tek® O.C.T. (Sakura, 4583), and cryosectioned at 12 μm, as described (Jo et al., 2017; Lakk et al., 2018). The sections were probed with antibodies against Piezo1 (Proteintech, 15939), α-SMA (Sigma, A2457), and Collagen IV (EMD Millipore, AB769). Secondary antibodies included anti-rabbit IgG DyLight 488 (Invitrogen, 35552), anti-mouse IgG DyLight 594 (Invitrogen, 35511), and anti-goat IgG Alexa 647 (Invitrogen, A21469). Sections were coverslipped with DAPI-Fluoromount-G (EMS, 17984-24) and imaged with Fluoview-1000 confocal microscope (Olympus, Center Valley, PA).

Yarishkin O., Phuong T.T., Baumann J.M., De Ieso M.L., Vazquez-Chona F., Rudzitis C.N., Sundberg C., Lakk M., Stamer W.D, & Križaj D. (2020). Piezo1 channels mediate trabecular meshwork mechanotransduction and promote aqueous fluid outflow. The Journal of physiology, 599(2), 571-592.

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Cell Cycle, Proliferation, and Apoptosis Analysis

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Flow cytometry was used to examine DAPI for cell cycle analysis, Ki-67 for cell proliferation, and cleaved caspase 3 expression for apoptosis following drug treatment. Cells were fixed with 4% formaldehyde and permeabilized using ice-cold methanol. The cells were then incubated with DAPI (10 mg/mL, 1:1000), mouse anti-human Ki-67 (Cell Signaling #9449, 1:400), and rabbit anti-human cleaved caspase 3 antibodies (Cell Signaling #9661, 1:800) at room temperature for 1 hour. Anti-mouse IgG DyLight 594 (Invitrogen #35511, 1:100) and anti-rabbit IgG DyLight 488 (Invitrogen #35553, 1:100) were used to detect anti-Ki-67 and anti-cleaved caspase 3 antibodies, respectively. The experiments were carried out on a CytoFLEX Flow Cytometry (Beckman Coulter, CA, USA), and the data was analyzed with FlowJo v10 from BD Biosciences (Franklin Lakes, NJ, USA).

Chen C.C., Wang S., Yang J.M, & Huang C.H. (2023). Targeting Ras signaling excitability in cancer cells through combined inhibition of FAK and PI3K. bioRxiv.

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TGFβ1-Induced EMT Signaling Pathway

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Recombinant human TGFβ1 was purchased from R&D Systems (USA). The proteasome inhibitor MG132 was purchased from Merck Millipore (USA). Cycloheximide (CHX), Methyl-β-cyclodextrin (MβCD) and TGFRI inhibitor (SB431542) was purchased from Sigma-Aldrich (USA). Trastuzumab, an anti-HER2 monoclonal antibody, was purchased from Roche (Switzerland). Antibodies against p-Akt, Akt, p-Smad2/3, Smad2/3, Smad4, β-actin, TGFRI (immunoprecipitation), TGFRII, and γ-catenin, as well as normal rabbit IgG, anti-rabbit, anti-mouse, and anti-rat IgG-HRP antibodies and protein A/G plus agarose, were obtained from Santa Cruz Biotechnology (USA). Anti-rabbit IgG-DyLight 488 was obtained from Thermo Fisher Scientific (USA). Antibodies against Caveolin-1, E-cadherin and N-cadherin were obtained from BD Transduction Laboratories (USA). An antibody against Vimentin was purchased from Thermo Scientific (USA). An antibody against Snail was purchased from Abcam Plc. (UK). An antibody against Slug was purchased from Cell Signaling Technology, Inc. (USA). An antibody against Smurf2, Smad7, Tollip, ubiquitin and TGFRI (immunoblotting) was purchased from GeneTex (USA).

Tsao S.M, & Hsu H.Y. (2016). Fucose-containing fraction of Ling-Zhi enhances lipid rafts-dependent ubiquitination of TGFβ receptor degradation and attenuates breast cancer tumorigenesis. Scientific Reports, 6, 36563.

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CELF2 Regulation of Autophagy in Colorectal Cancer Cells

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CRC lines HCT116, and SW480, obtained from ATCC (Manassas, VA), and were authenticated using a STR-based method. Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Corning, Corning, NY) with 10% heat-inactivated FBS (Sigma-Aldrich, St. Louis, MO) without antibiotics at 37 °C in the presence of 5% CO₂. Chloroquine diphosphate salt was obtained from Sigma Aldrich (St. Louis, MO). Primary antibodies targeting CELF2 (AV40324), and Atg12 (#4180), Beclin-1 (#4122), Atg5 (#12994) were obtained from Sigma-Aldrich (St. Louis, MO) and Cell Signaling (Danvers, MA), respectively. Secondary Anti-rabbit IgG Dylight 488 (#35553), used for immunofluorescence, and Hoescht 33342 nuclear counter stain were obtained from ThermoFisher (Waltham, MA).
In vitro knockdown of CELF2 was conducted using three pooled siRNA transcripts obtained from Santa Cruz Biotechnology, (CELF2: sc-44554; Control: sc-44236, Santa Cruz Biotechnology, Dallas, TX). The full-length coding region of human CELF2 was amplified by RT-PCR from HCT-116 cells and expressed as amino-terminal FLAG epitope-tagged proteins in plasmid pCMV-Tag2B (a cytomegalovirus immediate early promoter driven expression vector) after cloning at the HindIII and XhoI restriction sites.

New J., Subramaniam D., Ramalingam S., Enders J., Sayed A.A., Ponnurangam S., Standing D., Ramamoorthy P., O’Neil M., Dixon D.A., Saha S., Umar S., Gunewardena S., Jensen R.A., Thomas S.M, & Anant S. (2019). Pleotropic role of RNA binding protein CELF2 in Autophagy Induction. Molecular carcinogenesis, 58(8), 1400-1409.

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Western Blot Analysis of MAPK Signaling

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Whole-cell lysates were extracted using RIPA lysis buffer and a mixture of protease and phosphatase inhibitors (Minitab, Roche, Basel, Switzerland). Lysates were sonicated on ice, debris removed by centrifugation, and supernatants stored at −80 °C. Sodium dodecyl sulfate-polyacrylamide 12% gels were used to separate proteins, and proteins were transferred to nitrocellulose membranes. Membranes were blocked with Odyssey blocking buffer (Li-Cor, Lincoln, NE) in a 1:1 mixture with PBS 1% Tween-20 (PBST). Primary antibodies (phospho-MAPK, and total MAPK, Cell Signaling, Danvers, MA) were incubated overnight in 1:1 blocking buffer to PBST. Primary antibodies were detected using DyLight conjugated secondary antibodies (anti-rabbit IgG DyLight 488 (#35553), and anti-mouse IgG DyLight 800 (#35521) from Thermo Fisher (Waltham, MA)). Protein bands were detected using Li-Cor odyssey protein imaging system.

Kumar D., Yalamanchali S., New J., Parsel S., New N., Holcomb A., Gunewardena S., Tawfik O., Lominska C., Kimler B.F., Anant S., Kakarala K., Tsue T.T., Shnayder Y., Sykes K., Padhye S, & Thomas S.M. (2018). Development and characterization of an in vitro model for radiation-induced fibrosis. Radiation research, 189(3), 326-336.

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Quantitative Analysis of Metabolic Regulators

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ITGB2 (#73663), GluT4 (#2213), Akt (#4691), Phospho-Akt (#4060), PI3Kinase (#4292), Anti-rabbit IgG, HRP-linked Antibody (#7074), Anti-mouse IgG and HRP-linked Antibody (#7076) were obtained from Cell Signaling Technology. ITGB2 (ab53009), Phospho-PI3Kinase (ab182651) and a-SMA (ab5694) were obtained from the Abcam company. ITGB2 (10554-1-AP), HIF1a (66730-1-Ig), MCT1 (20139-1-AP), MCT4 (22787-1-AP), LDHB (14824-1-AP) and PDK4 (12949-1-AP) were obtained from the Proteintech company. ITGB2 (AF1730) was purchased from R&D. Secondary Anti-rabbit IgG Dylight 680 (#35568), Anti-rabbit IgG Dylight 488 (#35553) and Anti-mouse IgG Dylight 800 (#35521) were obtained from ThermoFisher.
The primer sequences used in this study were obtained from commercial sources and are displayed in Table S1.

Zhang X., Dong Y., Zhao M., Ding L., Yang X., Jing Y., Song Y., Chen S., Hu Q, & Ni Y. (2020). ITGB2-mediated metabolic switch in CAFs promotes OSCC proliferation by oxidation of NADH in mitochondrial oxidative phosphorylation system. Theranostics, 10(26), 12044-12059.

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