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Goat anti mouse alexa fluor 488

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Goat anti-mouse Alexa Fluor 488 is a secondary antibody conjugated with the Alexa Fluor 488 fluorescent dye. It is designed to bind to mouse primary antibodies and can be used in various immunochemical techniques that require fluorescent detection.

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Lab products found in correlation

Goat anti rabbit alexa fluor 594, by Abcam (6 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Alexa fluor 488 goat anti rabbit, by Abcam (3 mentions) Rabbit anti ki67, by Abcam (2 mentions) Mouse anti t2r7, by Santa Cruz Biotechnology (2 mentions) Formalin, by Merck Group (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Tween 20, by Merck Group (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Mouse anti human cd31, by Abcam (2 mentions) Alexa fluor 594 donkey anti rabbit, by Abcam (2 mentions) Mouse anti fbp2, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 633 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Alexa fluor 633 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Rabbit anti β actin, by Merck Group (1 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Rabbit anti hif 1α, by Bioss Antibodies (1 mentions) Mouse anti β actin, by Merck Group (1 mentions)

26 protocols using goat anti mouse alexa fluor 488

1

Immunofluorescence Staining of Cell Markers

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The immunofluorescence studies were performed as described previously [18 (link)]. The cells were fixed, permeabilized, and incubated overnight at 4 °C with the following respective primary antibodies: rabbit anti-Fbp2 (1:500, produced and tested as described previously [19 (link)]), mouse anti-Fbp2 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA, sc-271799), rabbit anti-Hif1α (1:500, Bioss Antibodies, Woburn, MA, USA, bs-0737R), rabbit anti-Ldha (1:500, Novus Biologials, Littleton, CO, USA, NBP1-48336), rabbit anti-Hk2 (1:500, Merck ab3279), rabbit anti-Ki67 (1:500, Abcam, Cambridge, UK, ab15580), mouse anti-αSMA (1:500, Merck a5228), mouse anti-β-actin (1:500, Sigma-Aldrich a1978), and rabbit anti-β-actin (1:500, Sigma–Aldrich a2066). The primary antibodies were detected using the following fluorophore-labelled secondary antibodies: goat anti-mouse-AlexaFluor488 (1:1000, Abcam, ab150113), goat anti-mouse-AlexaFluor633 (1:1000, Thermo Fisher Scientific, Waltham, MA, USA, a21050), goat anti-rabbit-AlexaFluor 488 (1:1000, Invitrogen a11034), and goat anti-rabbit-AlexaFluor633 (1:1000, Invitrogen a21070). Nuclei were counterstained with DAPI. In the controls, the primary antibodies were omitted.
Duda P., Janczara J., McCubrey J.A., Gizak A, & Rakus D. (2020). The Reverse Warburg Effect Is Associated with Fbp2-Dependent Hif1α Regulation in Cancer Cells Stimulated by Fibroblasts. Cells, 9(1), 205.
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2

Immunocytochemical Identification of T2R7 Receptors

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Receptors of the bitter membrane proteins (T2Rs) were also identified using the same immunocytochemistry procedure. Mouse anti-T2R7 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), were prepared in three dilutions (i.e., 1:100, 1:200 and 1:400) and a goat Anti-Mouse Alexa Fluor 488 (Abcam) secondary antibody was added as above stated.
Ftuwi H., Parri R, & Mohammed A.R. (2021). Novel, Fully Characterised Bovine Taste Bud Cells of Fungiform Papillae. Cells, 10(9), 2285.
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3

Bitter Taste Receptor 7 Localization

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There are more than 25 taste receptors, i.e., T2Rs, responsible for bitter taste transduction. BTBCs were stained with primary antibody raised against the extracellular regions T2R7 as stated above. Cells were labelled using three dilutions (i.e., 1:100, 1:200 and 1:400) of mouse anti-T2R7 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) using and a secondary antibody goat anti-mouse Alexa Fluor 488 (Abcam, Cambridge, UK).
Ftuwi H., Parri R, & Mohammed A.R. (2021). Novel, Fully Characterised Bovine Taste Bud Cells of Fungiform Papillae. Cells, 10(9), 2285.
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4

Immunofluorescence Staining of GPC-1

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All incubations were performed in a humidified chamber. Cells were fixed on the surface of dry glass slides, rehydrated in DPBS for 3 min, followed by blocking with 1% Bovine Serum Albumin (BSA) solution in DPBS for 1 h at room temperature. Cells were then incubated for 2 h at room temperature with an MIL-38 anti-GPC-1 mouse primary antibody diluted in blocking solution (1% BSA) at 10 μg/mL. At this step, one of the 4 slides was incubated with the blocking agent instead of the secondary antibody and was used as a secondary antibody alone control to monitor the non-specific binding of second antibody. Then, the glass slides were washed with DPBS 3 times for 5 min and air-dried. Then, cells were incubated for 1 h with goat-anti-mouse Alexa Fluor 488 (Abcam, Australia) diluted in the blocking solution at 4 μg/mL and washed with DPBS 3 times for 5 min after the incubation. Finally, cells were mounted with a Fluoroshield medium containing DAPI, covered with a coverslip, and observed under a fluorescence microscope.
Rzhevskiy A.S., Razavi Bazaz S., Ding L., Kapitannikova A., Sayyadi N., Campbell D., Walsh B., Gillatt D., Ebrahimi Warkiani M, & Zvyagin A.V. (2019). Rapid and Label-Free Isolation of Tumour Cells from the Urine of Patients with Localised Prostate Cancer Using Inertial Microfluidics. Cancers, 12(1), 81.
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5

Immunofluorescence Analysis of Nasal Cilia

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Nasal tissue, obtained by nasal brushing biopsy, was suspended in cell culture medium, spread onto glass slides, and then air dried. Samples were treated with 4% paraformaldehyde, 0.2% Triton-X 100 and 1% BSA before incubation with primary (overnight at 4 °C) and secondary (2 h at room temperature) antibodies. Rabbit polyclonal anti-DNAH5 (HPA037470, Sigma, Sweden) and mouse monoclonal anti-alpha tubulin (ab24610, Abcam, UK) antibodies were used at a dilution of 1:1000. Highly cross-adsorbed secondary antibodies, goat anti-mouse Alexa Fluor 488 (1:1000) and goat anti-rabbit Alexa Fluor 647 (1:1000), were obtained from Abcam. Confocal images were taken using a Leica TCS SP8 confocal laser scanning microscope (Leica, Jena, Germany).
Chen W., Guo Z., Li M., Sheng W, & Huang G. (2024). Next-Generation Sequencing-Based Copy Number Variation Analysis in Chinese Patients with Primary Ciliary Dyskinesia Revealed Novel DNAH5 Copy Number Variations. Phenomics, 4(1), 24-33.
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6

Immunofluorescence Staining of Smooth Muscle Cells

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Encapsulated cells were fixed for 30 min with 10% formalin (Sigma-Aldrich), permeabilized with 0.05% Tween20 (Sigma-Aldrich) in phosphate-buffered saline (PBS) (PBST), and blocked with 1% bovine serum albumin (BSA, Sigma-Aldrich) in PBST. Primary anti-mouse alpha smooth muscle actin (α-SMA, Abcam) was added 1:1000 overnight at 4 °C in blocking solution. Samples were washed with PBST, and secondary antibody goat-anti-mouse AlexaFluor 488 (Abcam, 1:300) in 1% BSA in PBST was added overnight at 4 °C. F-actin filaments were labeled with Rhodamine Phalloidin (ThermoFisher Scientific, 1:300) and the cell nucleus labeled with 4′,6-Diamidino-2-Phenylindole (DAPI, ThermoFisher Scientific, 1:1000).
Gonzalez Rodriguez A., Schroeder M.E., Walker C.J, & Anseth K.S. (2018). FGF-2 inhibits contractile properties of valvular interstitial cell myofibroblasts encapsulated in 3D MMP-degradable hydrogels. APL Bioengineering, 2(4), 046104.
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7

Virus Titer Determination by TCID50 Assay

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Titers of the virus stocks were determined by use of a 50% tissue culture infectious dose (TCID50) assay adapted for indirect detection of virus-infected cells. Briefly, tenfold viral dilutions were inoculated on Vero-E6 cells and incubated for 7 (DOBV, SEOV and TULV) or 12 days (PUUV) at 37°C, which corresponded to peak titers during virus culture (data not shown). Subsequently, cells were fixed using 4% PFA and permeabilized using 0.02% Triton X-100 in PBS. Subsequently, cells were washed with PBS and stained using mouse anti-hanta nucleoprotein (1:500; abcam, Cambridge, United Kingdom) (for PUUV, SEOV and TULV) or mouse anti-hantavirus Dobrava strain nucleoprotein (1:1,000; IBTsystems, Binzwangen, Germany) (for DOBV) as primary antibodies and goat anti-mouse-AlexaFluor488 (1:200; Abcam, Cambridge, United Kingdom) as a secondary antibody. Positive wells were visualized using a Leica DMIL LED fluorescence microscope and scored by eye. TCID50 titers were calculated using the Spearman and Kärber algorithm (Hierholzer and Killington, 1996 (link)).
Hoornweg T.E., Zutt I., de Vries A., Maas M., Hoogerwerf M.N., Avšič-Županc T., Korva M., Reimerink J.H, & Reusken C.B. (2020). Development of a Comparative European Orthohantavirus Microneutralization Assay With Multi- Species Validation and Evaluation in a Human Diagnostic Cohort. Frontiers in Cellular and Infection Microbiology, 10, 580478.
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8

Immunofluorescence Staining of Smooth Muscle Cells

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VIC-laden hydrogels were fixed at room temperature in 10% formalin (Sigma-Aldrich) for 30 min at room temperature and washed with phosphate-buffered saline (PBS, Thermo Fisher Scientific). Cells were then permeabilized for 1 hour using 0.1% Tritonx100 (Fisher Scientific) in PBS and blocked with 5% bovine serum albumin (BSA, Sigma-Aldrich) in PBS for 1 hour at room temperature. Primary mouse monoclonal anti-alpha smooth muscle actin (αSMA, Abcam) was added at 1:1000 in 5% BSA in PBS overnight at 4 °C. Samples were then washed with 0.05% Tween 20 (Sigma-Aldrich) in PBS, and secondary antibody goat-anti-mouse Alexa Fluor 488 (Abcam, 1:300), cytoplasm stain HCS CellMask Orange (Thermo Fisher Scientific, 1:5000) and nucleus label 4’,6-Diamidino-2-Phenylindole (DAPI, Thermo Fisher Scientific, 1:1000) in 5% BSA in PBS were added overnight at 4 °C.
Rodriguez A.G., Schroeder M.E., Grim J.C., Walker C.J., Speckl K.F., Weiss R.M, & Anseth K.S. (2021). Tumor Necrosis Factor-α promotes and exacerbates calcification in heart valve myofibroblast populations. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(3), e21382.
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9

Quantifying Neutrophil CFTR and LAMP1

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105 human neutrophils were seeded in coverslips coated with poly-L-lysine (Sigma Aldrich, St. Louis, MO). Cells were fixed, permeabilized and stained with mouse anti-human CFTR [CF3] (Abcam, Cambridge MA), wheat germ agglutinin Alexa Fluor 594 (Abcam, Cambridge MA), rabbit anti-human LAMP1 (Abcam, Cambridge MA), goat anti-mouse Alexa Fluor 488 (Abcam, Cambridge MA), goat anti-rabbit Alexa Fluor 594 (Abcam, Cambridge MA) and Hoechst 33342 (ThermoFischer scientific). Slides were imaged using apotome fluorescent microscope (Zeiss) and analyzed with ImageJ software (NIH).
Robledo-Avila F.H., Ruiz-Rosado J.D., Brockman K.L., Kopp B.T., Amer A.O., McCoy K., Bakaletz L.O, & Partida-Sanchez S. (2018). Dysregulated calcium homeostasis in cystic fibrosis neutrophils leads to deficient antimicrobial responses. Journal of immunology (Baltimore, Md. : 1950), 201(7), 2016-2027.
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10

Monocyte Attachment and Localization Assay

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Freshly-isolated PBM were incubated with polymyxin B and the indicated inhibitors for 45 minutes prior to ΔIgG stimulation at a density of 4×106 cells/mL. After 4 hours, 2×105 were seeded on poly-L-lysine coated coverslips for 15 minutes at 37 °C to allow attachment. Monocytes were fixed with 4% paraformaldehyde overnight at 4 °C, then blocked with 10% goat serum (Abcam, MA) for 1 hour at room temperature under permeabilized (0.1% Triton X-100) or non-permeabilized conditions. Cells were stained with mouse anti-human CD31 (Abcam, MA) overnight and goat anti-mouse Alexa Fluor 488 (Abcam, MA) plus wheat germ agglutinin (WGA) labeled with Texas red (ThermoFisher Scientific, IL) for 1 hour. Finally, coverslips were mounted with Fluoroshield solution with DAPI (Sigma-Aldrich, MO). Images were acquired using a Zeiss LSM800 confocal microscope, and then analyzed using ImageJ (NIH, USA).
Merchand-Reyes G., Robledo-Avila F.H., Buteyn N.J., Gautam S., Santhanam R., Fatehchand K., Mo X., Partida-Sanchez S., Butchar J.P, & Tridandapani S. (2019). CD31 acts as a checkpoint molecule and is modulated by FcγR-mediated signaling in monocytes. Journal of immunology (Baltimore, Md. : 1950), 203(12), 3216-3224.
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