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Alexa fluor 594conjugated goat anti mouse igg h l

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Alexa Fluor 594-conjugated goat anti-mouse IgG (H + L) is a secondary antibody that binds to mouse immunoglobulin (IgG) and its heavy and light chains. It is conjugated with the Alexa Fluor 594 fluorescent dye, which can be detected using appropriate instrumentation.

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Lab products found in correlation

Alexa fluor 488 conjugated goat anti rabbit igg h l, by Thermo Fisher Scientific (3 mentions) Primary antibodies for tnf α, by Proteintech (2 mentions) N storm a1, by Nikon (2 mentions) Antifade mounting medium with dapi, by Beyotime (2 mentions) Fv1000, by Olympus (2 mentions) Anti pax7, by Developmental Studies Hybridoma Bank (1 mentions) Affinipure fab fragment goat anti mouse igg, by Jackson ImmunoResearch (1 mentions) Alexa fluor 488 conjugated goat anti mouse igm, by Thermo Fisher Scientific (1 mentions) Anti ki67, by Cell Signaling Technology (1 mentions) Anti gapdh, by Abcam (1 mentions) Alexa fluor 488 conjugated goat anti mouse igm μ chain, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm, by BD (1 mentions) Facs fortessa, by BD (1 mentions) Pathway bioimaging system, by BD (1 mentions) Attovision software, by BD (1 mentions) Nigericin, by Merck Group (1 mentions) Sheep anti mouse igg hrp, by Cytiva (1 mentions) Anti human caspase 1, by Santa Cruz Biotechnology (1 mentions)

9 protocols using alexa fluor 594conjugated goat anti mouse igg h l

1

Immunofluorescence Staining of Muscle Stem Cells

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Antibodies used were anti-Pax7 (mouse IgG1, dilution 1:100, catalog No. AB_528428; Developmental Studies Hybridoma Bank, Iowa City, Iowa), antibody of laminin (rat or rabbit, dilution 1:1,500, catalog No. L0663 or L939; Sigma-Aldrich), anti-Ki67 (dilution 1:400, catalog No. 9129S; Cell Signaling Technology, Danvers, Massachusetts), anti-myogenin (F5D, dilution 1:100, sc-12732; Santa Cruz Bio-technology, Dallas, Texas), anti-GAPDH (dilution 1:2,500, ab9485; Abcam, Cambridge, Massachusetts), AffiniPure Fab fragment goat anti-mouse IgG (H + L, AFFGAI, 0.1 mg/ml; catalog No. 115–007-003; Jackson ImmunoResearch), AlexaFluor 594-conjugated goat anti-mouse IgG (H + L, dilution 1:1,500, catalog No. A-11032,; Life Technologies, Grand Island, New York), AlexaFluor 488-conjugated goat anti-mouse IgM (dilution 1:1,500, catalog No. A-21042; Life Technologies), and AlexaFluor 647-conjugated goat anti-rabbit or anti-mouse antibodies (dilution 1:1,500, catalog No. A-21244 or A-21235; Life Technologies). All reagents that are not specifically documented were purchased from Sigma-Aldrich.
YUE L., TALUKDER M.A., GURJAR A., LEE J.I., NOBLE M., DIRKSEN R.T., CHAKKALAKAL J, & ELFAR J.C. (2019). 4-AMINOPYRIDINE ATTENUATES MUSCLE ATROPHY AFTER SCIATIC NERVE CRUSH INJURY IN MICE. Muscle & nerve, 60(2), 192-201.
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2

Immunofluorescence Analysis of Lymphocytic Markers

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Maxillae with intact surrounding tissue were fixed in 4% paraformaldehyde, decalcified in Immunocal solution (Decal Chemical) for 14 d followed by placement in Cal-Arrest solution (Decal Chemical) to neutralize the pH of the tissue, and then embedded in OCT compound. Coronal sections (6-μm thick) were stained with mAbs to CD19 (6D5; Abcam), CD138 (EPR6454; Abcam), or BLyS (Buffy 2; Abcam) or polyclonal Ab to APRIL (Abcam), followed by appropriate secondary reagents conjugated to AlexaFluor594 or AlexaFluor488 (Life Technologies). Staining for Igs in tissues was performed using AlexaFluor488–conjugated goat anti-mouse IgM (μ chain) and AlexaFluor594–conjugated goat anti-mouse IgG (H+L) (Life Technologies).
The specificity of staining was confirmed using appropriate isotype control or non-immune IgG. Stained images were visualized and captured using a Nikon Eclipse Ni-E automated fluorescent microscope and NIS-Elements software.
Abe T., AlSarhan M., Benakanakere M.R., Maekawa T., Kinane D.F., Cancro M.P., Korostoff J.M, & Hajishengallis G. (2015). The B-cell stimulatory cytokines BLyS and APRIL are elevated in human periodontitis and are required for B-cell–dependent bone loss in experimental murine periodontitis. Journal of immunology (Baltimore, Md. : 1950), 195(4), 1427-1435.
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3

Cell Surface Receptor Immunophenotyping

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For cell surface staining, adherent cells (ARPE-19, MRC-9 and HAP1) were gently detached with a cell scraper and washed twice with PBS + 2% FBS, incubated with PDGFRα-specific mouse monoclonal antibody (BD Biosciences, clone αR1 and Sino biological, clone 5A10H9), mouse monoclonal anti-ErbB1 (Sigma, clone 225), mouse monoclonal anti-ErbB2, anti-ErbB3, anti-ErbB4 (R&D systems, Clone 191924, Clone 66223, Clone 182818) or mouse IgG1 anti-CMV pp72 (clone 6E1; Santa Cruz Biotechnology), mouse IgG2a H1P73, mouse IgG2b (MA5-14447; Invitrogen) at 2 μg/ml for 30 minutes on ice for isotype control. After two washes, cells were incubated with the following secondary antibodies: goat anti-Mouse IgG (H+L) AlexaFluor 594 conjugated (Invitrogen) at 2 μg/ml for 30 minutes on ice, washed twice and acquired with a FACSFortessa (BD Biosciences) flow cytometer. For intracellular staining, 293Expi cells were fixed and permeabilized with Cytofix/Cytoperm (BD Biosciences), according to the manufacturer’s instructions, and then stained with the anti-gO antibodies or 13H11 mab (as isotype control) at 2 μg/ml for 30 minutes on ice, washed and incubated with goat anti-Human IgG (H+L) AlexaFluor 594 conjugated (Invitrogen) at 2 μg/ml for 30 minutes on ice, washed twice and acquired with a FACSFortessa flow cytometer. Analysis was performed with FlowJo software.
Kabanova A., Marcandalli J., Zhou T., Bianchi S., Baxa U., Tsybovsky Y., Lilleri D., Silacci-Fregni C., Foglierini M., Fernandez-Rodriguez B.M., Druz A., Zhang B., Geiger R., Pagani M., Sallusto F., Kwong P.D., Corti D., Lanzavecchia A, & Perez L. (2016). Platelet-derived growth factor-α receptor is the cellular receptor for human cytomegalovirus gHgLgO trimer. Nature microbiology, 1(8), 16082.
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4

High-throughput Viral Infection Assay

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Confluent layers of ARPE-19, MRC-9 or HAP1 cells in 96-well flat-bottom plates were infected with a fixed amount of virus. MOI of infection (0.5) was chosen to give 500 to 1000 infected cells per microscopic field. 48 hours post-infection the cells were fixed with 5% acetic acid in methanol for 7 minutes on ice, washed four times with PBS and then stained with 0.5 μg/ml of mouse anti-pp72 antibody for 2 hours on ice. After four PBS washes, cells were incubated with 1 μg/ml of a goat anti-Mouse IgG (H+L) AlexaFluor 594 conjugated (Invitrogen). After four PBS washes, cells were counterstained with DAPI, images were acquired on BD Pathway bioimaging system and percentage of the infected cells was automatically calculated by BD AttoVision software. The number of infected cells was plotted as histograms and dose response curves were generated by plotting the relative infected cells against the ligand or antibodies dilutions.
Kabanova A., Marcandalli J., Zhou T., Bianchi S., Baxa U., Tsybovsky Y., Lilleri D., Silacci-Fregni C., Foglierini M., Fernandez-Rodriguez B.M., Druz A., Zhang B., Geiger R., Pagani M., Sallusto F., Kwong P.D., Corti D., Lanzavecchia A, & Perez L. (2016). Platelet-derived growth factor-α receptor is the cellular receptor for human cytomegalovirus gHgLgO trimer. Nature microbiology, 1(8), 16082.
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5

Inflammasome Activation Assay Protocol

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PMA (cat# P1585), ATP (cat# A7699), nigericin (cat# N7143), 4′,6-diamidino-2-phenylindole (DAPI; cat# D9542), and cytochalasin D (cat# C8273) were purchased from Sigma-Aldrich (St Louis, MO, USA), anti-ASC (cat# SC-22514-R), anti-human caspase-1 (cat# SC-56036), anti-mouse caspase-1 (cat# SC-514), anti-human IL-1β (cat# SC-32294), anti-α-tubulin (cat# SC-32293), anti-GAPDH (cat# SC-32233), and goat anti-rabbit IgG-horseradish peroxidase (HRP; cat# SC-2004) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-NLRP3 (cat# AG-20B-0014) from Adipogen (San Diego, CA, USA), anti-mouse IL-1β (cat# AF-401-NA) from R&D Systems Inc. (Minutesneapolis, MN, USA); sheep anti-mouse IgG-HRP (cat# NA931) from Amersham (Amersham, UK), (5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester (CFSE; cat# C1157) and Alexa Fluor-594 conjugated goat-anti-mouse IgG (H+L) (cat# A-11005) from Invitrogen (Carlsbad, CA, USA); and fluoresbrite yellow green carboxylate microspheres (1-μm YG beads, cat# 15702) from Polysciences Inc. (Warrington, PA, USA).
Chung I.C., OuYang C.N., Yuan S.N., Lin H.C., Huang K.Y., Wu P.S., Liu C.Y., Tsai K.J., Loi L.K., Chen Y.J., Chung A.K., Ojcius D.M., Chang Y.S, & Chen L.C. (2019). Pretreatment with a Heat-Killed Probiotic Modulates the NLRP3 Inflammasome and Attenuates Colitis-Associated Colorectal Cancer in Mice. Nutrients, 11(3), 516.
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6

Subcellular Localization of DSPP Mutants

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DSPP is not expressed in the HEK293 cells, so we choose this cell line to explore the location of the DSPP mutant. HEK293 cells were cultured in 24-well plates with Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Gibco®, USA), 100 unit/mL penicillin, and 100 μg/mL streptomycin. The cells were transfected with DSP-pEGFP-N1-WT or DSP-pEGFP-N1-MUT using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA). After 36 h of transfection, protein localization was detected by immunocytochemistry (ICC). The slides were incubated overnight at 4 °C with rabbit polyclonal anti-calnexin (CANX) antibody (cat: A0692, ABClonal, Wuhan, China) or mouse polyclonal anti-Golgi matrix protein (GM130) antibody (cat: ab169276, Abcam, Cambridge, UK) at a dilution of 1:600, followed by incubation with Alexa Fluor 594-conjugated goat anti-rabbit IgG(H+L) (cat: AS039, ABClonal, Wuhan, China) or Alexa Fluor 594-conjugated goat anti-mouse IgG(H+L) (cat:A-11005, Invitrogen, USA) for 2 h. Subsequently, the cells were incubated for 2 h with 4',6-diamidino-2-phenylindole (DAPI). Cells were imaged using a confocal microscope (Zeiss LSM 800, Jena, Germany).
Du Q., Cao L., Liu Y., Pang C., Wu S., Zheng L., Jiang W., Na X., Yu J., Wang S., Zhu X, & Yang J. (2021). Phenotype and molecular characterizations of a family with dentinogenesis imperfecta shields type II with a novel DSPP mutation. Annals of Translational Medicine, 9(22), 1672.
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7

Immunostaining and BrdU Assay Protocol

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Cells were fixed and permeabilized. Cells were blocked by 10% normal goat serum (Sigma-Aldrich, #G9023) in PBS for 1 h, then were incubated with primary antibodies for 1 h and with either Alexa Fluor™ 594-conjugated goat anti-mouse IgG (H + L; Invitrogen, #A11005, RRID:AB_2534073) or Alexa Fluor™ 488-conjugated goat anti-rabbit IgG (H + L; Invitrogen, #A11008, RRID:AB_143165) for 1 h. All immunostaining experiments in this study were accompanied with DAPI staining. The BrdU-incorporation assay was performed using the BrdU Labeling and Detection Kit I (Roche, #11296736001, RRID:AB_2814711) according to the manufacturer's instruction.
Ling H., Li Y., Peng C., Yang S, & Seto E. (2024). HDAC10 inhibition represses melanoma cell growth and BRAF inhibitor resistance via upregulating SPARC expression. NAR Cancer, 6(2), zcae018.
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8

Immunofluorescence Staining of Embryos

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The embryo was xed with 4% PFA for 15 min at room temperature and 0.3% Triton X-100 in PBS was added for 20 min to permeabilize the cell membrane. Then, the embryo was incubated with primary antibodies for TNF-α (1:100, Proteintech Inc., Rosemont, IL, USA) and CDX2 (1:100, Abcam, Inc., Cambridge, MA, USA) at 4 °C overnight. Subsequently, the embryo was incubated with Alexa Fluor 594conjugated goat anti-mouse IgG (H + L), (1:1000, Invitrogen, Inc., Carlsbad, CA, USA) and AlexaFluor 488conjugated goat anti-rabbit IgG (H + L) (1:1000, Invitrogen, Inc.) for 60 min at room temperature in the dark, followed by mounting with antifade mounting medium with DAPI (Beyotime, Inc., Shanghai, China).
The analysis was conducted using a confocal laser scanning microscope (Olympus FV1000, Olympus Corporation, Waltham, MA, USA) (Nikon N-STORM & A1, Nikon Corporation, Tokyo, Japan). The six embryos used for staining originated from the same volunteer.
Lv J., Shan X., Yang H., Wen Y., Zhang X., Chen H., Li H., Tian D., Wang C.C., Zhang R., Li T.C., Zhang X., Zhao X., Lu Y., Qin L., Zhu M, & Xu W. (2022). Single Cell Proteomics Profiling Reveals That Embryo-Secreted TNF-α Plays a Critical Role During Embryo Implantation to the Endometrium. Reproductive sciences (Thousand Oaks, Calif.), 29(5).
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9

Immunofluorescence Staining of Embryos

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The embryo was xed with 4% PFA for 15 min at room temperature and 0.3% Triton X-100 in PBS was added for 20 min to permeabilize the cell membrane. Then, the embryo was incubated with primary antibodies for TNF-α (1:100, Proteintech Inc., Rosemont, IL, USA) and CDX2 (1:100, Abcam, Inc., Cambridge, MA, USA) at 4 °C overnight. Subsequently, the embryo was incubated with Alexa Fluor 594conjugated goat anti-mouse IgG (H + L), (1:1000, Invitrogen, Inc., Carlsbad, CA, USA) and AlexaFluor 488conjugated goat anti-rabbit IgG (H + L) (1:1000, Invitrogen, Inc.) for 60 min at room temperature in the dark, followed by mounting with antifade mounting medium with DAPI (Beyotime, Inc., Shanghai, China).
The analysis was conducted using a confocal laser scanning microscope (Olympus FV1000, Olympus Corporation, Waltham, MA, USA) (Nikon N-STORM & A1, Nikon Corporation, Tokyo, Japan). The six embryos used for staining originated from the same volunteer.
Lv J., Shan X., Yang H., Wen Y., Zhang X., Chen H., Li H., Tian D., Wang C.C., Zhang R., Li T.C., Zhang X., Zhao X., Lü Y., Qin L., Zhu M., & Xu W. (2020). Single cell proteomics profiling reveals that embryo-secreted TNF-α plays a critical role during embryo implantation to the endometrium.
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