We used the following antibodies:anti-CD11b(1:500, ab8878, Abcam), anti-CD68(1:500, GB14043, Servicebio), anti-cleaved caspase-3 (Asp175) (5A1E) (1:400, #9664, Cell Signaling Technology, USA), anti-SARS-CoV-2 spike glycoprotein antibody-Coronavirus(1:500, ab272504, Abcam), anti-phospho Akt(1:800, 13038T, Cell Signaling Technology), anti-SQSTM1/p62(1:250, 23214S, Cell Signaling Technology), anti-BCL-2(1:200, sc-492, Santa Cruz Biotechnology), Alexa Fluor 647 goat anti-rat IgG(1:500, ab150159, Abcam), Alexa Fluor 488 goat anti-rabbit (1:500, ab150077, Abcam), Alexa Fluor 647 Goat anti-mouse (1:500, ab150115, Abcam), DAPI (Beyotime Institute of Biotechnology, China).
Alexa fluor 647 goat anti rat igg
Manufactured by Abcam
Sourced in United States
Alexa Fluor 647 goat anti-rat IgG is a secondary antibody conjugate that binds to rat immunoglobulin G (IgG) molecules. It is labeled with the Alexa Fluor 647 fluorescent dye, which can be detected using instruments that excite and detect in the near-infrared spectrum.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 goat anti rabbit igg, by Abcam (3 mentions)
Ab272504, by Abcam (1 mentions)
Ab150159, by Abcam (1 mentions)
Ab8878, by Abcam (1 mentions)
Gb14043, by Wuhan Servicebio Technology (1 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Anti cleaved caspase 3 asp175 5a1e, by Cell Signaling Technology (1 mentions)
Sc 492, by Santa Cruz Biotechnology (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Anti sqstm1 p62, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Ab150077, by Abcam (1 mentions)
Ab150115, by Abcam (1 mentions)
Ab228549, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Beyotime (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Antibody dilution buffer, by Solarbio (1 mentions)
Microscope slide, by Thermo Fisher Scientific (1 mentions)
Monoclonal rat anti cd3, by Abcam (1 mentions)
Confocal microscope, by Zeiss (1 mentions)
5 protocols using alexa fluor 647 goat anti rat igg
1
Multimarker Immunofluorescence for COVID-19 Analysis
2
Immunofluorescent Labeling of Amyloid-Beta
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Briefly, free-floating brain sections were treated with blocking buffer containing 5% bovine serum albumin (BSA, Beyotime, China) in PBS with 0.3% Triton X-100 (PBST) for one hour at 23 °C. Then brain sections were incubated with primary rabbit monoclonal antibodies for mouse anti-β amyloid (D54D2) XP® (1:1,000; 8243S; Cell Signaling Technology, US) diluted with 1% BSA in PBST overnight at 4 °C. After washing three times with PBS, sections were further incubated with secondary antibody (Alexa Fluor® 647 Goat Anti-Rat IgG, 1:1200; ab150167; Abcam, US) at 23 °C. The sections were then washed three times with PBS and counterstained with DAPI (5 μM in PBS; ab228549; Abcam, US), before being mounted with Prolong Gold antifade reagent (ThermoFisher; US).
3
Immunofluorescent Colon Tissue Staining
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Colon samples were embedded in O.C.T. (Sakura Finetek), cut in 4 μm sections, and adhered to microscope slides (Thermo Fisher Scientific). All slices were blocked with 100 μL of blocking solution for 30 min at room temperature in a humidified chamber. Then, the primary and secondary antibodies were incubated in a humidified chamber each at room temperature for 60 min. Phosphate-buffered saline (pH 7.4) was used to wash the slices following the primary and secondary antibody incubations three times for 5 min. And sections were counterstained with DAPI. The primary antibodies used were monoclonal rabbit anti-IL17A (Abcam) and monoclonal rat anti-CD3 (Abcam), respectively, and both were diluted to 1 : 200 in antibody dilution buffer (Solarbio). The primary antibodies were fluorescently labelled separately with Alexa Fluor 488 goat anti-rabbit IgG (Abcam) and Alexa Fluor 647 goat anti-rat IgG (Abcam) secondary antibodies, that were diluted to 1 : 1000 in antibody dilution buffer (Solarbio).
4
Quantifying Macrophage Phenotypes via Immunofluorescence
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RAW264.7 macrophages seeded on glass-bottom dishes were fixed in 4% paraformaldehyde (KeyGen Biotech, China) and then permeabilized with ice-cold 0.5% TritonX-100. The cells were blocked in PBS containing 10% bovine serum albumin (KeyGen Biotech, China) for 30 min and then were incubated with rabbit monoclonal antibody inducible nitric oxide synthase (iNOS) (Bioss Antibodies, China) and rat monoclonal antibody F4/80 (Abcam, USA) overnight at 4°C. After washing with PBS, cells were incubated with Alexa Fluor 488 goat anti-rabbit IgG (Abcam, USA) and Alexa Fluor 647 goat anti-rat IgG (Abcam, USA) for 1 h at 37°C. Finally, cells were counterstained with 50 mg/ml 4′,6-diamidino-2-phenylindole (DAPI) (KeyGen Biotech, China) before capturing images with a confocal microscope (Zeiss, Germany).
5
Immunofluorescence Staining of Mouse Tissues
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Immunofluorescence staining was performed as described previously.32 (link) Briefly, paraffin-embedded tissues of mice samples were cut into 3 µm sections and they were dewaxed in xylene and rehydrated in graded ethanol solutions. Antigen retrieval was performed with citrate buffer (6 pH) for 10 minutes at 95°C. Sections rinsed in PBS for 5 min. Non-specific labeling was blocked by incubation with 1% BSA at room temperature for 30 min. Sections were then incubated with anti-CD11c (1:200; Cell signaling Technology) and anti-F4/80 (1:200; Abcam) primary antibodies at 4°C overnight. Subsequently, Alexa Fluor 488 goat anti-rabbit IgG (1:200; Abcam) and Alexa Fluor 647 goat anti-rat IgG (1:200; Abcam) were used as secondary antibodies for 2 hours in the dark at room temperature. The nucleus was stained using SlowFade Diamond Antifade Mountant with DAPI (Life Technologies, Waltham, Massachusetts, USA). The samples were visualized by Keyence BZ-9000. All images were analyzed by Keyence BZ-X Analyzer software.
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