Alexa fluor 594 antibodies

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor 594 antibodies are fluorescent labeling reagents designed for use in various immunological techniques, such as flow cytometry, immunohistochemistry, and Western blotting. These antibodies are conjugated with the Alexa Fluor 594 dye, which has an excitation maximum of 590 nm and an emission maximum of 617 nm.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Lsm 710, by Zeiss (2 mentions) Fv1000 confocal laser scanning microscopy, by Olympus (1 mentions) Alexa fluor 488 antibody, by Thermo Fisher Scientific (1 mentions) Anti opsin antibody, by Merck Group (1 mentions) Anti ha antibody, by Roche (1 mentions) Ab157107, by Abcam (1 mentions) Upright fluorescent microscope, by Leica (1 mentions) Ab11197, by Abcam (1 mentions) Ab76126, by Abcam (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Premier coverslips, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Dm4000b, by Leica (1 mentions)

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5 protocols using alexa fluor 594 antibodies

Immunofluorescence Assay for LC3 and TFEB

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LO2 cells seeded in a 12-well plate were fixed with 4% paraformaldehyde for 15 min at RT, washed with PBS three times, and permeabilized with frozen methanol for 10 min at −20 °C. After blocking with 5% BSA for 30 min, cells were incubated with primary antibodies targeting LC3 (1:250) or TFEB (1:50, 13372-1-AP, Proteintech, Chicago, IL, USA) in NCM Universal Antibody Diluent (New Cell & Molecular Biotech, Suzhou, China) at 4 °C overnight after being washed for three times. Next, cells were incubated with anti-rabbit Alexa Fluor 594 antibodies (1:100, Thermo Fisher Scientific, Waltham, MA, USA) in 5% BSA at RT for 1 h and stained with DAPI. Olympus FV1000 Confocal Laser Scanning Microscopy was used to observe the cells.

Chen J., Zheng D., Cai Z., Zhong B., Zhang H., Pan Z., Ling X., Han Y., Meng J., Li H., Chen X., Zhang H, & Liu L. (2023). Increased DNMT1 Involvement in the Activation of LO2 Cell Death Induced by Silver Nanoparticles via Promoting TFEB-Dependent Autophagy. Toxics, 11(9), 751.

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Opsin and Cpf1 Localization in Retina

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For the analysis of the opsin-positive area, formalin-fixed paraffin embedded samples were prepared at day 42 post-injection (n = 4). Cross-section samples were immunostained with anti-opsin antibody (Millipore, cat. no. AB5405, 1:1000) and Alexa Fluor 488 antibody (Thermo Fisher Scientific, cat. no. A-11034, 1:500). The opsin-positive area was measured using Image J software (1.47 v, NIH) by blinded observers. To visualize the distribution of HA-tagged Cpf1, the eyes were fixed in 4% paraformaldehyde for 1 h at room temperature. RPE complexes (RPE/choroid/sclera) were prepared for immunostaining and then incubated with anti-HA antibody (Roche, cat. no. 3F10, 1:100) overnight at 4 °C. After staining with Alexa Fluor 594 antibodies (Thermo Fisher Scientific, cat. no. A-11006, 1:500), the RPE flat-mounts were imaged using a confocal microscope (LSM 710, Carl Zeiss). The scanning parameters were as follows: scaling (x = 0.042 μm/pixel, y = 0.042 μm/pixel, z = 0.603 μm/pixel), dimensions (x = 1024, y = 1024, channels: 2, 8-bit) with objective C-Apochromat 40 × /1.20 W Korr M27. ZEN 2 software was used to process the images.

Koo T., Park S.W., Jo D.H., Kim D., Kim J.H., Cho H.Y., Kim J., Kim J.H, & Kim J.S. (2018). CRISPR-LbCpf1 prevents choroidal neovascularization in a mouse model of age-related macular degeneration. Nature Communications, 9, 1855.

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Immunohistochemical Characterization of Tumor Organoids

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On day 7, HGA and LGA tumor organoids were washed twice with DPBS and fixed with 4% paraformaldehyde for 1 h and then additionally washed twice with DPBS. Cultures were histologically processed, paraffin embedded, and sectioned at 5-μm thickness. Tissue sections on microscope slides were then stained with hemotoxylin and eosin (H&E).
IHC was used to visualize biomarkers mucin 2 (MUC2) and cytokeratin 20 (CK20). Blocking was performed by incubation under Dako Protein Block for 15 min. Primary antibodies MUC2 (ab11197, abcam, raised in mouse), CK20 (ab76126, abcam, raised in rabbit), or CD44 (ab157107, abcam, raised in rabbit) were applied to the sections on the slides at a 1:200 dilution in Dako Antibody Diluent and incubated at room temperature for 1 h. Next, secondary Alexa Fluor 488 or Alexa Fluor 594 antibodies (ThermoFisher) with appropriate species reactivity were applied to all samples at 1:200 in Dako Antibody Diluent and left at room temperature for 1 h (anti-mouse Alexa Fluor 488 and anti-rabbit Alexa Fluor 594, Life Technologies, Carlsbad, CA, A-11070). Sections were then incubated with Dapi for 5 min before coverslipping. Sections were imaged on a Leica upright fluorescent microscope.

Votanopoulos K.I., Mazzocchi A., Sivakumar H., Forsythe S., Aleman J., Levine E.A, & Skardal A. (2018). Appendiceal Cancer Patient-Specific Tumor Organoid Model for Predicting Chemotherapy Efficacy Prior to Initiation of Treatment: A Feasibility Study. Annals of surgical oncology, 26(1), 139-147.

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Immunofluorescence Analysis of β-Catenin

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Immunofluorescence analysis of β-catenin was performed in cells seeded in 8-well chamber slides (Lab-Tek-II), fixed in formaldehyde/PBS and incubated in a methanol/acetone solution for 15 minutes. Cells were incubated with anti-β-catenin (1:500 dilution), washed in PBS and then probed with AlexaFluor 594 antibodies (Life Technologies). Nuclei were counterstained with 0.1 mg/ml 4',6-diamidino-2-phenylindole (DAPI, Life Technologies), as described previously [36 (link)].

Maftouh M., Belo A.I., Avan A., Funel N., Peters G.J., Giovannetti E, & van Die I. (2014). Galectin-4 expression is associated with reduced lymph node metastasis and modulation of Wnt/β-catenin signalling in pancreatic adenocarcinoma. Oncotarget, 5(14), 5335-5349.

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Immunohistochemical Analysis of Corneal Collagen

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Eight-micron thick corneal sections were prepared, postfixed at room temperature for 10 min, and blocked with 5% donkey serum for 1 h at room temperature. Immunostaining was performed using collagen I (COL1) (1:200; Abcam), kept overnight at 4°C, and secondary stained with Alexa-Fluor 594 antibodies (1:500; Life Technologies) for 4 h. A drop of DAPI antifade Vectashield medium (H1200) was applied, and sections were mounted with premier coverslips (Thermo Fisher). (DAPI; Vector Laboratories) The stained sections were viewed and photographed with a fluorescence microscope (Leica DM 4000B, Leica Microsystems Inc., Buffalo Grove, IL) equipped with a digital camera (SpotCam RT KE, Diagnostic Instruments Inc., Sterling Heights, MI). Negative control samples were stained by excluding primary or secondary antibody in serially sectioned slides.

Sinha N.R., Balne P.K., Bunyak F., Hofmann A.C., Lim R.R., Mohan R.R, & Chaurasia S.S. (2021). Collagen matrix perturbations in corneal stroma of Ossabaw mini pigs with type 2 diabetes. Molecular Vision, 27, 666-678.

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