Alexa fluor 594 conjugated anti rabbit

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

Alexa Fluor 594-conjugated anti-rabbit is a fluorescent-labeled secondary antibody. It is designed to detect and visualize rabbit primary antibodies in various immunodetection techniques, such as immunofluorescence microscopy, flow cytometry, and Western blotting.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

19 protocols using alexa fluor 594 conjugated anti rabbit

Immunofluorescence Staining of EGFR and TNFR1

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

For immunofluorescence staining, HT-29 and HeLa cells were cultured on coverslips in 12-well plates. Post treatment with the indicated reagents, the cells were fixed with 4% paraformaldehyde for 30 min at RT. Anti-EGFR (Cell Signalling, 4267S) (1:50) and anti-TNFR1 (R&D systems, AD225) (1:50) or Anti-HA (Santa Cruz, sc805) (1:50) and anti-FLAG (Sigma Aldrich, F3165) (1:50) were incorporated to each sample in immunofluorescence blocking buffer (PBS with 3% BSA, 1% saponin and 1% Triton X-100) and then incubated at 4 °C overnight. Each sample was washed thrice with PBS. Alexa Fluor 488-conjugated anti-goat (Invitrogen, A-10055) (1:100) and Alexa Fluor 594-conjugated anti-rabbit (Invitrogen, A-21207) (1:100) or Alexa Fluor 488-conjugated anti-mouse (Invitrogen, A-11029) (1:100) and Alexa Fluor 594-conjugated anti-rabbit antibodies (Invitrogen, A-21207) (1:100) were incorporated into each sample in immunofluorescence blocking buffer and incubated at RT for 1 h. The nuclei were stained with DAPI for 5 min and evaluated using LSM980 (Carl Zeiss, Oberkochen, Germany) and ZEN (blue edition) programme.

Nam Y.W., Shin J.H., Kim S., Hwang C.H., Lee C.S., Hwang G., Kim H.R., Roe J.S, & Song J. (2024). EGFR inhibits TNF-α-mediated pathway by phosphorylating TNFR1 at tyrosine 360 and 401. Cell death and differentiation.

+ Open protocol

+ Expand

Multiplexed Immunofluorescence Staining of Mouse Tumor Tissue

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

Paraffin mouse tumor tissue sections were pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 30 min, and blocked with normal goat serum (Cat# AR0009, Boster, China) for 1 h at room temperature. Then, the slides were performed with rat anti-mouse IDO1 (1:100, Cat# 122402, BioLegend), and rabbit anti-mouse CD8 antibody (1:200, Cat# ab203035, Abcam) overnight at 4 °C. After the slides were washed with PBS, they were incubated with Alexa Fluor 633-conjugated anti-rat (1:200, Cat# A-21094, Invitrogen) or Alexa Fluor 594-conjugated anti-rabbit (1:200, Cat# R37117, Life Technologies) secondary antibodies for 2 h in the dark at room temperature. These antibodies were used for labeling the rat anti-IDO1 antibody or the rabbit anti-CD8 antibody, respectively. Slides were mounted using VECTASHIELD antifade mounting medium containing 4,6-diamidino-2-phenylindole (DAPI) (Cat# H-1200, Vector Laboratories) according to manufacturer’s recommendations. Fluorescent staining was visualized under a laser scanning confocal microscope (TCS SP8, Leica, Germany).

Lou Q., Liu R., Yang X., Li W., Huang L., Wei L., Tan H., Xiang N., Chan K., Chen J, & Liu H. (2019). miR-448 targets IDO1 and regulates CD8+ T cell response in human colon cancer. Journal for Immunotherapy of Cancer, 7, 210.

+ Open protocol

+ Expand

Immunostaining of Cardiomyocytes

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

NRVMs were prepared for staining by means of Cardiomyocyte Immunocytochemistry Kit (Thermo Fisher Scientific) following manufacturer’s instruction. Briefly, NRVMs were fixed with Fixative Solution for 30 min, permeabilized with Permeabilization Solution S for 15 min at room temperature and then blocked with Blocking Solution for 30 min. Mitochondria were stained using anti-TOM20 antibody (Santa Cruz; 1:500, rabbit) and sarcomeres were stained using anti-α-sarcomeric actinin antibody (Sigma; 1:500, mouse) over-night at 4°C. The day after, samples were incubated with Alexa Fluor 594 conjugated anti-rabbit (Life Technologies, 1:250) and Alexa Fluor 488 conjugated anti-mouse (Life Technologies, 1:250) for 1h at room temperature. Coverslips were mounted using NucBlue™ Fixed Cell ReadyProbes™ Reagent with DAPI to stain nuclei (Invitrogen). Images were collected at Zeiss LSM 700 confocal system equipped with a PlanApo 40x/1.2 oil objective, and an Argon laser used to excite the fluorophores at the appropriate wavelength. Images were collected at 2048×2048 pixels per image with a 100 Hz acquisition rate. Images were analyzed using the Fiji distribution of the Java-based image processing program ImageJ [38 (link)]. The experiments were performed both in normoxia and after 6h of anoxia followed by 30 min of reoxygenation.

Antonucci S., Di Sante M., Sileikyte J., Deveraux J., Bauer T., Bround M.J., Menabò R., Paillard M., Alanova P., Carraro M., Ovize M., Molkentin J.D., Cohen M., Forte M.A., Bernardi P., Di Lisa F, & Murphy E. (2019). A novel class of cardioprotective small-molecule PTP inhibitors. Pharmacological research, 151, 104548.

+ Open protocol

+ Expand

Fluorescent Embryo Imaging with IF Staining

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

Transgenic fluorescent embryos were embedded in 1% agarose in a glass‐bottom dish. Immunofluorescence double staining was performed with chicken‐anti‐GFP (1:400; Life Technologies) and rabbit‐anti‐phospho‐histone3 (pH3) antibodies (1:250; Abcam). We used AlexaFluor488‐conjugated anti‐chicken (1:1,000; Life Technologies) and AlexaFluor594‐conjugated anti‐rabbit (1:1,000; Life Technologies) secondary antibodies to reveal primary antibodies. Confocal imaging was performed using a Nikon inverted A1r spectral.

Cacialli P., Mailhe M., Wagner I., Merkler D., Golub R, & Bertrand J.Y. (2022). Synergistic prostaglandin E synthesis by myeloid and endothelial cells promotes fetal hematopoietic stem cell expansion in vertebrates. The EMBO Journal, 41(19), e108536.

+ Open protocol

+ Expand

Phospho-Histone H3 Immunofluorescence Assay

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

The phospho-histone H3 immunofluorescence assay was performed as previously described [49 (link)] with primary anti-phoshpo-histone H3 (Cell Signaling #3377) and secondary Alexa Fluor 594 conjugated anti-rabbit (Life Technologies #A-11037) antibodies. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Images were acquired using a Nikon Eclipse Ti microscope with NIS Elements imaging software (Nikon, Melville, NY).

Shepherd J.H., Uray I.P., Mazumdar A., Tsimelzon A., Savage M., Hilsenbeck S.G, & Brown P.H. (2016). The SOX11 transcription factor is a critical regulator of basal-like breast cancer growth, invasion, and basal-like gene expression. Oncotarget, 7(11), 13106-13121.

+ Open protocol

+ Expand

Imaging DNA Damage in Transgenic Embryos

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

Transgenic fluorescent embryos were embedded in 1% agarose dissolved in 1xE3 in a glass-bottom dish. For immunofluorescence, double staining was performed using chicken-anti-GFP (1:500; Life Technologies), rabbit-anti-H2A.X (phospho Ser139) (1:500; GTX127342; Genetex) rabbit, and pH3 (1:250; Abcam) antibodies. We used AlexaFluor488-conjugated anti-chicken (1:1,000; Life Technologies) and AlexaFluor594-conjugated anti-rabbit (1:1,000; Life Technologies) secondary antibodies to reveal primary antibodies. Confocal imaging was performed using a Nikon inverted A1r spectral.

Cacialli P., Dogan S., Linnerz T., Pasche C, & Bertrand J.Y. (2023). Minichromosome maintenance protein 10 (mcm10) regulates hematopoietic stem cell emergence in the zebrafish embryo. Stem Cell Reports, 18(7), 1534-1546.

+ Open protocol

+ Expand

Multimodal Immunofluorescence of Protein Markers in Tissue Samples

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

Similar to immunohistochemistry, tissue epitopes were unmasked and subsequently exposed to UV light for 24 h to reduce autofluorescence. Sections were incubated with blocking buffer for 2 h and with primary antibodies (tau, Aβ and GFAP) for 72 h at 4 °C twice. In addition, for staining with HSP90AA1, HSP90AB1 and BAG3 antibodies, the sections were incubated with blocking buffer for 1 h and with the antibodies for 48 h or 24 h at 4 °C (for details, see Supplementary Materials Table S1).
Subsequently, the sections were incubated with Alexa Fluor 594-conjugated anti-rabbit, Alexa Fluor 488-conjugated anti-mouse or Alexa Fluor 647-conjugated anti-goat antibodies (1:200; Thermo Fisher, Waltham, Massachusetts, USA) for 2 h and then with 0.05% DAPI for 10 min at room temperature. Sections were mounted and coverslipped with PVA-DABCO.

Gonzalez-Rodriguez M., Villar-Conde S., Astillero-Lopez V., Villanueva-Anguita P., Ubeda-Banon I., Flores-Cuadrado A., Martinez-Marcos A, & Saiz-Sanchez D. (2021). Neurodegeneration and Astrogliosis in the Human CA1 Hippocampal Subfield Are Related to hsp90ab1 and bag3 in Alzheimer’s Disease. International Journal of Molecular Sciences, 23(1), 165.

+ Open protocol

+ Expand

Hypoxia and Proliferation Regulation

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

PCL (M_n = 700,000–900,000), sodium orthovanadate, sodium fluoride, β-glycerophosphate, DOX (doxorubicin), and BrdU were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pimonidazole hydrochloride was obtained from Hypoxyprobe Inc. (Burlington, MA, USA). The following antibodies were used for the study: Alexa Fluor 594 phalloidin (Thermo Fisher Scientific, Waltham, MA, USA), anti-HIF-1α (R&D Systems, Minneapolis, MN, USA), anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA), anti-F-actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti-BrdU (Thermo Fisher Scientific, Waltham, MA, USA), anti-pimonidazole (Hypoxyprobe Inc.), anti-Ki67 (Cell Signaling Technology, Beverly, MA, USA), and cleaved caspase 3 (Cell Signaling Technology, Beverly, MA, USA). The following secondary antibodies were used: Alexa Fluor 488-conjugated anti-mouse (Thermo Fisher Scientific, Waltham, MA, USA), Alexa Fluor 594-conjugated anti-mouse (Thermo Fisher Scientific, Waltham, MA, USA), Alexa Fluor 594-conjugated anti-rabbit (Thermo Fisher Scientific, Waltham, MA, USA), and HRP-conjugated anti-mouse (Cell Signaling Technology, Beverly, MA, USA).

Oh E.T., Kim H.G., Choi M.H., Lee J.S., Kim S.J., Kwak J.Y, & Park H.J. (2021). Multi-Layer Nanofibrous PCL Scaffold-Based Colon Cancer Cell Cultures to Mimic Hypoxic Tumor Microenvironment for Bioassay. Cancers, 13(14), 3550.

+ Open protocol

+ Expand

Subcellular Localization Analysis of Proteins

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

HeLa cells were cultured on coverslips in 24-well plates overnight. After transient transfections, cells were washed with PBS, fixed with 4% paraformaldehyde–PBS for 20 min at RT, washed with PBS, permeabilized with 0.3% Triton X-100 in PBS for 15 min and washed again with PBS. After being blocked with a solution containing 2% BSA, 0.05% Triton X-100 in PBS for 30 min at room temperature (RT), cells were incubated with anti-COX IV and anti-Flag antibodies in blocking solution for 1 h at RT. Upon washing with blocking solution, bound antibodies were subsequently detected by incubation, as appropriate, AlexaFluor 594-conjugated anti-rabbit (11,012, Invitrogen) and AlexaFluor 488-conjugated anti-mouse (A11001, Invitrogen) secondary antibodies in blocking solution for 1 h at RT. After staining with 1 mg/mL DAPI in PBS, cells were examined by using a Leica confocal microscope.

Boutoual R., Jo H., Heckenbach I., Tiwari R., Kasler H., Lerner C.A., Shah S., Schilling B., Calvanese V., Rardin M.J., Scheibye-Knudsen M, & Verdin E. (2022). A novel splice variant of Elp3/Kat9 regulates mitochondrial tRNA modification and function. Scientific Reports, 12, 14804.

+ Open protocol

+ Expand

DNA Damage Response Protein Immunoblotting

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

Antibodies used were as follows: anti-CtIP rabbit polyclonal (1/1000; Bethyl, USA), anti-Mre11 mouse monoclonal (1/1000; GeneTex, USA), anti-NBS1 mouse monoclonal (1/ 1000; GeneTex), anti-actin mouse monoclonal (1/5000; Sigma-Aldrich), anti-Rad51 mouse polyclonal (1/500; Abnova, Taiwan), anti-Rad51 rabbit polyclonal (1/1000; Bio Academia, Japan), anti-BrdU mouse monoclonal (1/100; BD Pharmingen, USA), anti-BrdU rat polyclonal (1/250; Abcam, UK), anti-cyclin A rabbit polyclonal (1/100; Santa Cruz Biotechnology); Alexa Fluor 488-conjugated anti-mice IgG (1/1000; Molecular Probes), Alexa Flour 594-conjugated anti-mice IgG (1/1000; Molecular Probes), Alexa Fluor 594-conjugated anti-rabbit (1/1000; Invitrogen) and Alexa Flour 488-conjugated anti-rabbit IgG (1/1000; Life Technologies).

Hoa N.N., Akagawa R., Yamasaki T., Hirota K., Sasa K., Natsume T., Kobayashi J., Sakuma T., Yamamoto T., Komatsu K., Kanemaki M.T., Pommier Y., Takeda S, & Sasanuma H. (2015). Relative contribution of four nucleases, CtIP, Dna2, Exo1 and Mre11, to the initial step of DNA double-strand break repair by homologous recombination in both the chicken DT40 and human TK6 cell lines. Genes to cells : devoted to molecular & cellular mechanisms, 20(12), 1059-1076.

+ 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!