Mouse tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Sections were stained with hematoxylin and eosin or oil red according to standard protocols. Immunohistochemical staining of paraffin sections was carried out with 1:50 anti-UCP-1 (Abcam), 1:50 p-CREB (Ser133) (CST), and 1:50 PKA C (CST) and detected by Inverted microscope (Olympus). The cell sizes and areas of adipose tissues were measured by Image J. Immunofluorescence staining of paraffin sections for BrdU incorporation was carried out with primary antibodies 1:50 anti-UCP-1 (Abcam), 1:100 BrdU (Santa Cruz), or 1:200 CD68 (Bio-Rad, Formerly AbD Serotec) and secondary antibodies Alexa Fluor 594 conjugated goat antibody to rabbit IgG, Alexa Fluor 488 conjugated goat antibody to mouse IgG, or Alexa Fluor 594 conjugated goat antibody to rat IgG (Invitrogen, 1:1,000). The Elecron microscopic observations were conducted through scanning electron microscope (PHILIPS CM120). All the representative images were repeated in at least 3 independent experiments.
Alexa fluor 594 conjugated goat antibody to rabbit igg
Manufactured by Thermo Fisher Scientific
Alexa Fluor 594 conjugated goat antibody to rabbit IgG is a secondary antibody reagent. It is used to detect and visualize rabbit primary antibodies in various immunoassay techniques.
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Lab products found in correlation
Anti ucp1, by Abcam (3 mentions)
Inverted microscope, by Olympus (1 mentions)
Bromodeoxyuridine (brdu), by Abcam (1 mentions)
Cm120, by Philips (1 mentions)
Bromodeoxyuridine (brdu), by Santa Cruz Biotechnology (1 mentions)
Beta catenin antibody, by Cell Signaling Technology (1 mentions)
Streptavidin pe cyanine7, by Thermo Fisher Scientific (1 mentions)
Mito tracker green, by Beyotime (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Click it plus edu alexa fluor 488 imaging kit, by Thermo Fisher Scientific (1 mentions)
Anti pdgfrα, by R&D Systems (1 mentions)
Anti red fluorescent protein, by Rockland Immunochemicals (1 mentions)
Anti dpp4, by R&D Systems (1 mentions)
Affinipure fab fragment, by Jackson ImmunoResearch (1 mentions)
4 protocols using alexa fluor 594 conjugated goat antibody to rabbit igg
1
Histological and Immunohistochemical Analysis of Mouse Adipose Tissue
2
Profiling Innate Immune Cells by Flow Cytometry
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For flow cytometry, the following fluorochrome-conjugated anti-mouse antibodies were used: CD45 (30-F11), CD11b (M1/70), CD3e (145-2C11), CD4 (GK1.5), F4/80 (BM8), Foxp3 (FJK-16s), CD127 (A7R34), KLRG1 (2F1) were from eBioscience; IL-33R-biotin (T1/S2, clone DJ8) from MD Bioproducts; Streptavidin PE-Cyanine7 was from eBioscience; FITC anti-mouse Lineage Cocktail with Isotype Ctrl was from Biolegend. For immunofluorescence staining, purified anti-mouse IL-33 polyclonal antibody (AF3626) and anti-human IL-33 polyclonal antibody (AF3625) were from R&D Systems; beta-Catenin antibody (Cat: #9587) was from Cell Signaling Technology and Pan-keratin antibody (PCK-26) was from Abcam. Alexa Fluor 488-conjugated Donkey antibody to mouse IgG (A21202), Alexa Fluor 594-conjugated Donkey antibody to goat IgG (A11058) and Alexa Fluor 594-conjugated goat antibody to rabbit IgG (A11037) were from Invitrogen.
3
Immunofluorescence Imaging of Mitochondria and Adipocyte Markers
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The mitochondria were stained by Mito-Tracker Green (Beyotime) for 30 min in DMEM. Cells were fixed with 4% paraformaldehyde and incubated with 1:50 anti-UCP-1 (Abcam) or 1:100 anti-PAK C (CST) or 1:100 anti-Perilipin (CST) antibody and further detected by secondary antibodies Alexa Fluor 594 conjugated goat antibody to rabbit IgG (Invitrogen 1:1,000). Cells were then washed in PBS and stained with 40, DAPI. Images were acquired by fluorescence microscopy (Zeiss System) or High Content Screening Microplate Imaging Reader (Thermo Fisher Scientific). The representative images were repeated in at least 3 independent experiments.
4
Multimodal Immunohistochemistry and Fluorescence Imaging
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Mouse tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Sections were stained with hematoxylin and eosin according to standard protocols. Immunohistochemical staining of paraffin sections was carried out with 1:200 anti-UCP1 (Abcam). For Immunofluorescence, slides were fixed with 4% PFA and permeabilized in ice cold methanol. Heat-mediated antigen retrieval with a 0.01 M citric acid (pH 6.0) was performed for 5 min in a microwave. After blocking with 4% BSA, the sections were blocked with unconjugated AffiniPure Fab Fragment (1:100; Jackson), then incubated with primary antibodies anti-UCP1 (rabbit; 1:200; Abcam), anti-red fluorescent protein (rabbit; 1:500; Rockland), anti-PDGFRα (goat; 1:200; R&D) or anti-DPP4 (goat; 1:200; R&D) followed by detection with secondary antibodies Alexa Fluor® 594 conjugated goat antibody to rabbit IgG (1:1,000; Invitrogen) or Alexa Fluor® 647 conjugated donkey antibody to goat IgG (1:1,000; Invitrogen). Finally, the sections were incubated with DAPI or Hoechst. The adipocytes were stained with BODIPY® 493/503 (Invitrogen). EdU incorporation assay was performed following the instruction of Click-iT® Plus EdU Alexa Fluor® 488 Imaging Kit (Invitrogen).
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