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Alexa fluor 488 labeled goat anti mouse

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Alexa Fluor®488-labeled goat anti-mouse is a fluorescently-labeled secondary antibody. It is designed for use in immunofluorescence applications to detect and visualize target proteins labeled with mouse primary antibodies.

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Lab products found in correlation

Alexa fluor 568 labeled goat anti rabbit, by Thermo Fisher Scientific (3 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 labeled goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Axio observer d1 microscope, by Zeiss (2 mentions) Halotag tmr ligand, by Promega (2 mentions) Alpha plan apochromat 100 1.40 oil immersion objective, by Zeiss (2 mentions) Brightline single band laser filter set, by IDEX Corporation (2 mentions) Axio observer microscope, by Zeiss (1 mentions) Rabbit anti gfap, by Merck Group (1 mentions) Alexa fluor 568 labeled goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Mouse anti map2, by Santa Cruz Biotechnology (1 mentions) Nucblue live cell stain readyprobes, by Thermo Fisher Scientific (1 mentions) Ti e inverted microscope, by Oxford Instruments (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Mouse anti lamp1, by Abcam (1 mentions) Alexa fluor 568, by Thermo Fisher Scientific (1 mentions) Application suite, by Leica (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) α sma, by Merck Group (1 mentions) Cisplatin, by Merck Group (1 mentions)

Spelling variants of this product

Alexa Fluor 488 goat anti-mouse (726 citations) Anti-mouse Alexa Fluor 488 (195 citations) Anti-mouse Alexa-488 (181 citations) Goat anti-mouse Alexa 488 (164 citations) Alexa Fluor 488 goat anti (9 citations) Alexa Fluor 488 anti (6 citations) Anti-mouse Alexa Fluor (4 citations) Alexa-488 anti-goat (3 citations) Anti-mouse Alexa Fluor 488-labeled (2 citations) Fluor 488 labeled anti-mouse (2 citations)

9 protocols using alexa fluor 488 labeled goat anti mouse

1

Neuronal Cell Culture Purity and Morphology

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The purity of mouse primary neuronal cell cultures as well the morphological changes in neurons after 48 h of treatment with lactacystin (2.5 μg/ml) and celastrol (1 μM) was determined by immunocytochemistry. Cells were seeded in 24-well plates containing culture glass cover slips covered by poly-ornithine (0.01 mg/ml) at a density of 3 × 105 cells/well. After cell treatment at 7 DIV, the cultures were fixed with 4 % paraformaldehyde, permeabilized with PBS containing 0.25 % Triton X-100 (PBS-TX-100) and blocked with 5 % normal goat serum in PBS-TX-100. Primary antibodies against neuronal (mouse anti-MAP-2, 1:200; Santa Cruz) and glia (rabbit anti-GFAP, 1:400; Sigma) markers were added and incubated with cells for 120 min at RT. After thrice washing in PBS, the cells were incubated for 60 min with secondary antibodies: Alexa Fluor®488-labeled goat anti-mouse and Alexa Fluor®568-labeled goat anti-rabbit IgG (Invitrogen, USA) diluted 1:500 in PBS. After washing with PBS, cover slips with cells were mounted with ProLong®Gold antifade reagent (Invitrogen, USA). Cells were examined using a fluorescence AxioObserver microscope (Carl Zeiss, Germany) equipped with the software Axiovision 3.1 at excitation wavelengths of 470 nm (Alexa Fluor®488) and 555 (Alexa Fluor®568).
Konieczny J., Jantas D., Lenda T., Domin H., Czarnecka A., Kuter K., Śmiałowska M., Lasoń W, & Lorenc-Koci E. (2014). Lack of Neuroprotective Effect of Celastrol Under Conditions of Proteasome Inhibition by Lactacystin in In Vitro and In Vivo Studies: Implications for Parkinson’s Disease. Neurotoxicity Research, 26(3), 255-273.
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2

Immunofluorescent Staining of HepaRG Cells

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HepaRG cells were rinsed in phosphate-buffered saline (PBS), fixed in 4% paraformaldehyde in PBS for 20 minutes, and quenched with 10 mM glycine in PBS. Cells were permeabilized for 5 minutes in 0.2% Triton-X100/PBS, followed by a 30-minute blocking step with 1% bovine serum albumin (BSA) in PBS. Cells were then incubated for overnight at 4°C with the following primary antibodies: rabbit anti-ZIP14 (Sigma 1:200), mouse anti-MRP2 (Abcam, 1:200), mouse anti-Lamp1 (Abcam, 1:100), mouse anti-beta1 NaK-ATPase (Abcam, 1:200, followed by a 45 minutes incubation with secondary antibodies AlexaFluor568-labeled goat anti-rabbit (Molecular Probes, 1:500), AlexaFluor568-labeled goat anti-mouse (Molecular Probes, 1:500), AlexaFluor488-labeled goat anti-rabbit (Invitrogen, 1:500), or AlexaFluor488-labeled goat anti-mouse (Invitrogen, 1:500). Inserts were mounted using ProLong Diamond Antifade Mountant (Invitrogen). Prior to fixation, nuclei were stained with NucBlue Live Cell Stain ReadyProbes according to manufacturer’s instructions (Invitrogen). HepaRG cells were imaged with a Yokogawa CSU-X1 spinning disk confocal system with a Nikon Ti-E inverted microscope using a 60x or 100x Plan Apo objective lens with Zyla cMOS camera using 405, 561 and 488 lasers. NIS elements software was utilized for acquisition parameters, shutters, filter positions and focus control.
Thompson K.J, & Wessling-Resnick M. (2019). ZIP14 is degraded in response to manganese exposure. Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine, 32(6), 829-843.
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3

Fluorescent Labeling of Actin and Vinculin

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Cells were fixed for 7 minutes in 3.7% paraformaldehyde and permeabilized for 4 minutes in 0.1% Triton X-100. Actin was detected using Phalloidin coupled to Alexa-Fluor 568 (Invitrogen, Carlsbad, CA). Vinculin was detected using the monoclonal antibody hVIN1 (ab11194, Abcam, Cambridge, UK). The secondary antibody used was Alexa-Fluor-488-labeled goat anti-mouse (Invitrogen, Carlsbad, CA). Coverslips were mounted using Fluoromount G (SouthernBiotech, Birmingham, AL) as an anti-fading reagent.
Images of fixes samples were acquired with a confocal microscope (Leica SP5) equipped with a 63× oil objective. Images were processed using the Leica Application Suite (Leica, Wetzlar, Germany) and ImageJ software.
Hind L.E., MacKay J.L., Cox D, & Hammer D.A. (2014). Two-Dimensional Motility of a Macrophage Cell Line on Microcontact-Printed Fibronectin. Cytoskeleton (Hoboken, N.J.), 71(9), 542-554.
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4

Analyzing Tumor Markers in Cell Lines

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Epirubicin (EPI) and Cisplatin (CS) were purchased from Sigma-Aldrich and dissolved in water and DMSO, respectively. Phosphoramide mustard (PM), the active metabolite of Cyclophosphamide, from National Cancer Institute and dissolved in DMSO. Human recombinant LH was a gift from Merck KGaA (Germany).
The following primary antibodies were used: Tubulin (T9026, Sigma-Aldrich); GADPH (G9545, Sigma-Aldrich); LHCGR (sc-25828, Santa Cruz); FOXL2 (ab5096, Abcam); γH2AX (05-636, Millipore); pan-cytokeratin (pan-CK, Z0622, Dako); αSMA (A2547, Sigma-Aldrich); RAD51 (sc-8349, Santa Cruz Biotechnology); ERCC1 (sc-17809, Santa Cruz Biotechnology); MLH1 (sc-271978, Santa Cruz Biotechnology). HRP-conjugated secondary antibodies were as follows: donkey anti-goat (Jackson ImmunoResearch) (1:50000); goat anti-mouse (GE Healthcare) (1:5000); goat anti-rabbit (GE Healthcare) (1:5000). The following secondary antibodies were used for IF analysis: Cy3-labeled donkey anti-goat antibody (1:400); Alexa Fluor 488-labeled goat anti-mouse (Thermo Fisher Scientific) (1:500); Alexa Fluor 568-labeled goat anti-mouse (Thermo Fisher Scientific) (1:500); Alexa Fluor 488-labeled goat anti-rabbit (Thermo Fisher Scientific) (1:500).
Marcozzi S., Rossi V., Salvatore G., Di Rella F., De Felici M, & Klinger F.G. (2019). Distinct effects of epirubicin, cisplatin and cyclophosphamide on ovarian somatic cells of prepuberal ovaries. Aging (Albany NY), 11(22), 10532-10556.
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5

Immunocytochemical Analysis of SH-SY5Y Cells

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Either differentiated or undifferentiated SH-SY5Y cells (0.5 × 106 cells) were cultured overnight on BioCoat poly-D-lysine/laminin coverslips (Corning) and subsequently fixed for 15 min at room temperature using 4% paraformaldehyde in phosphate-buffered saline (PBS) (Affymetrix). Coverslips were washed five times with PBS-T (PBS containing 0.1% Triton X-100), treated with 3% bovine serum albumin in TBS-T (PBS containing 0.1% Tween20) for 30 min at room temperature, washed once with TBS-T, and incubated in primary antibody diluted in TBS-T overnight at 4 °C. Coverslips were then washed extensively with TBS-T and then incubated with 1:1,000 (v/v) Alexa Fluor 488-labeled goat anti-mouse or Alexa Fluor 594-labeled goat anti-rabbit IgG (Thermo Fisher Scientific) and 1 μg/ml 4',6-Diamidine-2'-phenylindole dihydrochloride (DAPI) for 2 h at room temperature. After further extensive washing, coverslips were mounted using ProLong Gold antifade reagent (Thermo Fisher Scientific). Images were captured by an Olympus FV-1000 confocal laser scanning microscope.
Kurosaki T., Mitsutomi S., Hewko A., Akimitsu N, & Maquat L.E. (2022). Integrative Omics Indicate FMRP Sequesters mRNA From Translation and Deadenylation In Human Neuronal Cells. Molecular cell, 82(23), 4564-4581.e11.
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6

Immunofluorescent Detection of GFP in COS-7 Cells

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COS-7 cells were maintained 37 °C in 5% CO2 incubators. Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and 1% Pen-Strep was used as culture media. For immunofluorescent detection of GFP, cells were cultured on 4 well chamber slides. Cells were transfected using Lipofectamine LTX with Plus Reagent (Thermo Fisher, MA) using manufacturer’s protocol. 48 h after transfection, cells were fixed with 4% paraformaldehyde for 15 min, washed with 1x PBS, and permeabilized 0.1% triton X-100. Cells were then blocked with 5% normal goat serum in 1x PBS containing 0.1% triton X-100 for 1 h and incubated with GFP (1:1000) plus GA (1:100), GP (1:500), or GR (1:500) antibodies overnight at 4 °C. Slides were washed with 1x PBS and incubated with AlexaFluor 488 labeled goat anti-mouse (Thermo Fisher, MA; 1:500) and Alexa Fluor 555 labeled goat anti-rabbit (Thermo Fisher, MA; 1:500) antibodies and visualized with an Olympus epifluorescence microscope with Slidebook 5.5 software with identical fluorescent settings for each slide.
He F., Flores B.N., Krans A., Frazer M., Natla S., Niraula S., Adefioye O., Barmada S.J, & Todd P.K. (2020). The carboxyl termini of RAN translated GGGGCC nucleotide repeat expansions modulate toxicity in models of ALS/FTD. Acta Neuropathologica Communications, 8, 122.
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7

Immunoblotting and Immunofluorescence for HCV

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Primary antibodies used were against HCV core (Institute of Immunology, Tokyo, Japan), HCV NS5A (Virogen, Watertown, MA), β-Actin (Abcam, Cambridge, UK), Nrf2 (Santa Cruz, Dallas, TX), and p-Nrf2, phospho-serine 40, (Abcam). For immunoblot analysis, HRP-labeled secondary antibodies against mouse IgG, rabbit IgG, and goat IgG (GE Healthcare) were used depending on the primary antibodies used for immunoblotting. For immunofluorescence staining, Alexa-fluor-488-labeled goat anti-mouse and Alexa-fluor-568-labeled goat anti-rabbit (Life Technologies) secondary antibodies were used.
Murakami Y., Sugiyama K., Ebinuma H., Nakamoto N., Ojiro K., Chu P.S., Taniki N., Saito Y., Teratani T., Koda Y., Suzuki T., Saito K., Fukasawa M., Ikeda M., Kato N., Kanai T, & Saito H. (2018). Dual effects of the Nrf2 inhibitor for inhibition of hepatitis C virus and hepatic cancer cells. BMC Cancer, 18, 680.
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8

Confocal Microscopy of Condensate Formation

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An Axio Observer D1 Microscope (Zeiss, Germany) equipped with an Alpha Plan-Apochromat 100 × /1.40 Oil immersion Objective (Zeiss, Germany) and an Evolve 512 × 512 EMCCD camera with pixel size 16.0 μm (Photometrics; Tucson, AZ) was used for in vitro condensate formation, immunofluorescence, and live-cell imaging. A Brightline® single-band laser filter set (Semrock; excitation filter: FF01–561/14, emission filter: FF01–609/54, and dichroic mirror: Di02-R561–25 × 36) was used for the excitation and emission of HaloTag® TMR ligand (Promega; G8251) or Alexa Fluor 568-labeled goat anti-rabbit (Life Technologies; A11011). A Brightline® single-band laser filter set (Semrock; excitation filter: FF02–482/18–25, emission filter: FF01–525/45–25, and dichroic mirror: Di02-R488–25 × 36) was used for the excitation and emission of Alexa Fluor 488-labeled goat anti-mouse (Life Technologies; A11029). The microscope and EMCCD camera were controlled by the computer via SlideBook 6.0 software (3i). Images were processed and presented using Adobe PhotoShop (Adobe Inc).
Kent S., Brown K., Yang C.H., Alsaihati N., Tian C., Wang H, & Ren X. (2020). Phase-Separated Transcriptional Condensates Accelerate Target-Search Process Revealed by Live-Cell Single-Molecule Imaging. Cell reports, 33(2), 108248.
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9

Confocal Microscopy of Condensate Formation

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An Axio Observer D1 Microscope (Zeiss, Germany) equipped with an Alpha Plan-Apochromat 100 × /1.40 Oil immersion Objective (Zeiss, Germany) and an Evolve 512 × 512 EMCCD camera with pixel size 16.0 μm (Photometrics; Tucson, AZ) was used for in vitro condensate formation, immunofluorescence, and live-cell imaging. A Brightline® single-band laser filter set (Semrock; excitation filter: FF01–561/14, emission filter: FF01–609/54, and dichroic mirror: Di02-R561–25 × 36) was used for the excitation and emission of HaloTag® TMR ligand (Promega; G8251) or Alexa Fluor 568-labeled goat anti-rabbit (Life Technologies; A11011). A Brightline® single-band laser filter set (Semrock; excitation filter: FF02–482/18–25, emission filter: FF01–525/45–25, and dichroic mirror: Di02-R488–25 × 36) was used for the excitation and emission of Alexa Fluor 488-labeled goat anti-mouse (Life Technologies; A11029). The microscope and EMCCD camera were controlled by the computer via SlideBook 6.0 software (3i). Images were processed and presented using Adobe PhotoShop (Adobe Inc).
Kent S., Brown K., Yang C.H., Alsaihati N., Tian C., Wang H, & Ren X. (2020). Phase-Separated Transcriptional Condensates Accelerate Target-Search Process Revealed by Live-Cell Single-Molecule Imaging. Cell reports, 33(2), 108248.
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