Cells were grown on 35-mm cell culture dishes with glass bottoms (NEST, Wuxi, China). Cells were fixed with 4% paraformaldehyde for 30 min. Fixed cells were washed and permeabilized with 0.1% Triton X-100 for 10 min. Cells were blocked with goat serum for 30 min and incubated with the anti-β-catenin antibody (1:100, 8480S, CST) overnight at 4 °C. After washing with PBS, the cells were subsequently incubated with Alexa Fluor® 488-conjugated anti-rabbit IgG antibody (1:1000, ab150077, Abcam) for 1 h. The nuclei were stained with DAPI (62248, Thermo Scientific). Images were acquired using a Zeiss LSM710 confocal microscope (Zeiss, Germany) at 400× magnification.
Alexa fluor 488 conjugated anti rabbit igg antibody
Manufactured by Abcam
Sourced in United Kingdom
Alexa Fluor 488-conjugated anti-rabbit IgG antibody is a secondary antibody that binds to rabbit primary antibodies. It is conjugated to the Alexa Fluor 488 fluorescent dye, which emits green fluorescence when excited with the appropriate wavelength of light. This antibody can be used for detecting and visualizing target proteins in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions)
Alexa fluor 594 conjugated anti mouse igg antibody, by Abcam (2 mentions)
35 mm cell culture dishes, by NEST Biotechnology (1 mentions)
Glass bottoms, by NEST Biotechnology (1 mentions)
Anti caspase 1 antibody, by Abcam (1 mentions)
Alexa fluor 568 conjugated anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions)
Anti rank antibody, by Abcam (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 conjugated anti rabbit igg antibody, by Abcam (1 mentions)
Alexa fluor 488 conjugated anti mouse igg antibody, by Abcam (1 mentions)
Anti cd41, by Abcam (1 mentions)
Anti nlrp3, by Abcam (1 mentions)
Triton x, by Merck Group (1 mentions)
Blocking one histo, by Nacalai Tesque (1 mentions)
Fv1200, by Olympus (1 mentions)
Mouse anti cgrp, by Abcam (1 mentions)
Rabbit anti laminin, by Abcam (1 mentions)
Dylight 649, by Vector Laboratories (1 mentions)
5 protocols using alexa fluor 488 conjugated anti rabbit igg antibody
1
Immunofluorescence Staining of β-Catenin
2
Immunocytochemistry for Macrophage Markers
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Stimulated and unstimulated macrophages were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X‐100, and stained with a single antibody or combinations of anti–gasdermin D antibody (no. LS‐B4537; LifeSpan Biosciences), anti–caspase 1 antibody (no. ab‐1872; Abcam), anti–lipin 2 antibody (no. HPA017857; Sigma‐Aldrich), anti‐RANK antibody (no. ab222215; Abcam), and anti–IL‐1R1 antibody (no. ab106278; Abcam) for 12 hours, followed by Alexa Fluor 568–conjugated anti‐mouse IgG antibody (Invitrogen) or Alexa Fluor 488–conjugated anti‐rabbit IgG antibody (Abcam). DAPI (Molecular Probes) was used to visualize nuclei. Signals were visualized with a confocal laser scanning microscope (Leica SP8). Image processing was performed with Imaris 9.2.1 software.
3
Immunofluorescence Analysis of Inflammasome Components
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Platelets or PMNs were fixed and incubated with antibodies including anti-NLRP3 (Abcam, 1:100, catalog no. ab4207), anti-ASC (Novusbio, 1:100, catalog no. NBP1-78978), anti-CD41 (Abcam, 1:200, catalog no. ab134131), Alexa Fluor 488 anti-mouse CD41 (BioLegend, 1:200, catalog no. 133908), Hoechst (Thermo Fisher Scientific, 1:3,000, catalog no. H3570), anti-myeloperoxidase (Genetx, 1:200, catalog no. gtx75318), anti-myeloperoxidase (Abcam, 1:200, catalog no. ab208670) and anti-histone H3 (citrulline R2 + R8 + R17, 1:200, Abcam, catalog no. ab5103) at 4 °C overnight. Secondary antibodies included Alexa Fluor 647-conjugated anti-goat antibody (Abcam, 1:200, catalog no. ab150135), Alexa Fluor 647-conjugated anti-rabbit IgG antibody (Abcam, 1:200, catalog no. ab150079), Alexa Fluor 594-conjugated anti-rabbit IgG antibody (Abcam, 1:200, catalog no. ab150084), Alexa Fluor 594-conjugated anti-mouse IgG antibody (Abcam, 1:200, catalog no. ab150116), Alexa Fluor 488-conjugated anti-goat IgG antibody (BioLegend, 1:200, catalog no. 405508), Alexa Fluor 488-conjugated anti-rabbit IgG antibody (Abcam, 1:200, catalog no. ab150077) and Alexa Fluor 488-conjugated anti-mouse IgG antibody (Abcam, 1:200, catalog no. ab150113). Cells were captured using immunofluorescence confocal microscopy (×63 oil immersion lens, Leica SP8).
4
Immunofluorescence Analysis of Cryogenic Tissue Sections
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The sampled fetus or placenta tissues were frozen in OTC compound in cryomolds. On the other hand, sampled skeletal muscles were fixed on a corkboard with tragacanth gum and then frozen in isopentane at the temperature of liquid nitrogen. 8-μm-thick cryo-tissue sections were prepared using a cryostat, fixed with acetone for 5 min, and subjected to immunofluorescence staining as follows. After washing with PBS, tissue sections were incubated with 0.5% Triton-X (Sigma-Aldrich) in PBS for 10 min for permeabilization, washed with PBS, blocked using Blocking One Histo (NACALAI TESQUE, Kyoto, Japan) for 15 min, incubated with primary antibodies (rabbit anti-GFP and mouse anti-dystrophin, 1:100 diluted, Abcam, Cambridge, UK), washed, and incubated with Alexa-Fluor 488-conjugated anti-rabbit IgG antibody (1:200 diluted, Abcam) and Alexa-Fluor 588-conjugated anti-mouse IgG antibody. The immunostained tissue sections were observed under a confocal laser scanning fluorescence microscope (FV1200, Olympus, Tokyo, Japan) and Cell Sens Standard software (FV1-ASW, Olympus).
5
Immunolabeling of Spinal Cord and DRG Tissues
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One month after dorsal root crush injury, rats were euthanized with pentobarbital sodium and perfused with 4% PFA. Under a stereomicroscope, spinal cords with attached roots and DRG tissues were collected from perfused rats. Subsequently, tissues were postfixed in 4% PFA for 24 h, and cryoprotected in 30% sucrose (wt/vol) for at least 48 h at 4 °C. The tissues were then embedded in OCT (Sakura, Finetek, USA), cryosectioned using a cryostat, and mounted onto slides. Spinal cord and DRG tissues were cryosectioned at thicknesses of 25 µm and 15 μm, respectively. After blocking in 5% BSA with 0.1% Triton X-100 for 1 h, sections were incubated with primary antibodies at 4 °C overnight. Secondary antibodies were applied for 2 h at room temperature. The following primary antibodies were used: mouse anti-CGRP (1:1000; Abcam); rabbit anti-laminin (1:2000; Abcam); and IB4 conjugated with DyLight 649 (1:500; Vector). The corresponding secondary antibodies were used as follows: Alexa Fluor 488-conjugated anti-rabbit IgG antibody (1:1000; Abcam) and Alexa Fluor 594-conjugated anti-mouse IgG antibody (1:1000; Abcam). Images were acquired by confocal microscopy (Zeiss 710) and analyzed using ImageJ software.
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