Frozen adipose sections were prepared as above and incubated with the primary antibody mixture of GLUT4 (ab654, Abcam, United States), CD86 (ab119857, Abcam, United States), or CD206 (ab8918, Abcam, United States) at 4°C overnight. Then slides were protected from light and incubated with secondary Alexa Fluor 488- or Alexa Fluor 555-labeled goat anti-rabbit or mouse antibodies (A-11034, A-21422, Molecular Probes, United States) for 2 h at room temperature. Counterstaining was followed with DAPI for 15 min in an aluminum foil-covered box. Finally, the stained tissues were examined under a fluorescence microscope (Leica, Germany). Six mice for each group were used, and more than eight fields were chosen and analyzed for each adipose section. The fluorescence index was determined with Image Pro Plus 6.0 (Media Cybernetics, United States). The differences in fluorescence are displayed as a relative percent normalized to the respective control group (SC-PBS group in Figure 2 , SC group in Figure 3 , OHF-Sed-PBS group in Figure 4 , and OHF-Sed group in Figure 5 ).
Ab119857
Manufactured by Abcam
Sourced in United States, United Kingdom
Ab119857 is a recombinant monoclonal antibody that recognizes the human Cytochrome P450 1A1 protein. The antibody is suitable for use in various immunodetection applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab64693, by Abcam (3 mentions)
Ab1793, by Abcam (2 mentions)
Ab16667, by Abcam (2 mentions)
Af2535, by R&D Systems (2 mentions)
Ab178945, by Abcam (2 mentions)
Ab183685, by Abcam (2 mentions)
Ab654, by Abcam (1 mentions)
Ab8918, by Abcam (1 mentions)
A11034, by Thermo Fisher Scientific (1 mentions)
A21422, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Leica camera (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Ab83053, by Abcam (1 mentions)
Ab28696, by Abcam (1 mentions)
Ab92726, by Abcam (1 mentions)
Nb600 633, by Novus Biologicals (1 mentions)
Sc 18355, by Santa Cruz Biotechnology (1 mentions)
Ab53444, by Abcam (1 mentions)
Ab8878, by Abcam (1 mentions)
Alexa fluor 555 secondary antibody, by Thermo Fisher Scientific (1 mentions)
12 protocols using ab119857
1
Adipose Tissue Immunofluorescence Analysis
2
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protein concentration was determined by bicinchoninic acid protein assay. Equal amount of protein was used, and Western blotting was performed as previously described (66 (link)). Primary antibodies included anti-CD9 antibody (ab92726, Abcam, MA), anti-CD63 antibody (216130, Abcam, MA), anti-CD81 antibody [10037, Cell Signaling Technology (CST)], anti–annexin A1 antibody (3299, CST), anti-HSP90 antibody (4877, CST), anti–IL-6R antibody (ab83053, Abcam, MA), anti-Stat3 antibody and anti-p-Stat3 antibody (9139S and 9145S, CST), anti–IL-6 antibody (12912, CST), anti–nuclear factor κB antibody (8242S, CST), anti–TNF-α antibody (ab1793, Abcam, MA), anti–IL-1b antibody (NB600-633, Novus), anti-CD86 antibody (ab119857, Abcam, MA), anti-Arg1 antibody (sc-18355, Santa Cruz Biotechnology, CA), and anti–β-actin antibody (ab28696, Abcam, MA). The gray intensity of protein blots was measured using ImageJ software (NIH).
3
Corneal Immunohistochemistry Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cornea flat mounts, cryosections, and cultured cells were fixed in 4% paraformaldehyde and blocked with 0.5% Triton X-100/5% bovine serum albumin for 2 hours at room temperature. The samples were incubated with primary antibodies overnight at 4°C and then with secondary antibodies for 1 hour. The primary antibodies included anti-CD11b antibody (ab8878, Abcam), anti-CD68 antibody (ab53444, Abcam), anti-CD86 antibody (ab119857, Abcam), anti-CD206 antibody (AF2535, R&D Systems), and anti-Ki67 antibody (ab16667, Abcam). Secondary antibodies included donkey anti-rabbit immunoglobulin G (IgG) (H + L) Alexa Fluor 555, donkey anti-rat IgG (H + L) Alexa Fluor 488, donkey anti-rabbit IgG (H + L) Alexa Fluor 488, and donkey anti-goat IgG (H + L) Alexa Fluor 555 secondary antibodies (1:1000; Invitrogen). TUNEL staining (In Situ Cell Death Detection Kit, Fluorescein; Roche, IN, USA) was performed according to the manufacturer’s instructions. The corneal whole mounts were dissected into four petals and detected by ZEISS (LSM880, Germany). Three images of each petal were captured in mid-periphery areas of each cornea, respectively.
4
Immunofluorescence Staining of Mouse Heart and Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies used for immunofluorescence double-staining or triple-staining of mice heart tissue sections in each group are as follows: anti-CD86 antibody (ab119857, Abcam, USA), anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-NLRP3 antibody (ab214185, Abcam, USA), anti-ASC-antibody (sc-514414, Santa Cruz, USA), anti-Caspase-1 antibody (ab1872, Abcam, USA), Alexa Fluor 488 (D001-34, Abcam, USA). DAPI (c1002, Beyotime, China) stained cell nucleus. The brief process is as follows: dewaxing, rehydration, repair, blocking, primary antibody incubation at 4°C overnight, PBS washing, secondary antibody incubation in the dark, DAPI staining for cell nuclei, and mounting.
RAW264.7 macrophages were seeded in confocal dishes, after stimulated administration, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% TritonX-100 for 20 min. Then, cells were blocked with 1% BSA in the 37°C incubator for 1 h. Incubation with primary antibody at 4°C overnight, followed by incubation with the secondary antibody for 1 h at room temperature away from light. Then, wheat germ agglutinin (WGA) (AC15L012, Life-iLab, China), labeled with AF488, stained cell membrane for 20 min. Finally, nuclei were stained with DAPI for 5 min. The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).
RAW264.7 macrophages were seeded in confocal dishes, after stimulated administration, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% TritonX-100 for 20 min. Then, cells were blocked with 1% BSA in the 37°C incubator for 1 h. Incubation with primary antibody at 4°C overnight, followed by incubation with the secondary antibody for 1 h at room temperature away from light. Then, wheat germ agglutinin (WGA) (AC15L012, Life-iLab, China), labeled with AF488, stained cell membrane for 20 min. Finally, nuclei were stained with DAPI for 5 min. The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).
5
Histological Analysis of Mouse Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were euthanized via cervical dislocation, following which small intestine, colon, iWAT, epididymal WAT (eWAT), and BAT samples were immediately collected. Samples were then quickly placed in a freezer at −80 °C until use. Subsequently, the samples were harvested and fixed in 4% formaldehyde overnight for histological examination.
Paraffin-embedded small intestine, colon, and iWAT sections (5 μm) were stained with hematoxylin and eosin (H&E) or periodic acid Schiff (PAS) [14 (link)]. For immunohistochemical (IHC) staining, the primary IHC antibodies against UCP1 (ab10983, 1:1000), CD86 (ab119857, 1:1000), and ZO-1 (abG041, 1:1000) were purchased from Abcam (Cambridge, UK). Images of tissue samples were acquired using an Olympus BX53 microscope (Tokyo, Japan). Image-Pro Plus v.6.0 software was used to analyze adipocyte diameters in the H&E-stained sections.
Paraffin-embedded small intestine, colon, and iWAT sections (5 μm) were stained with hematoxylin and eosin (H&E) or periodic acid Schiff (PAS) [14 (link)]. For immunohistochemical (IHC) staining, the primary IHC antibodies against UCP1 (ab10983, 1:1000), CD86 (ab119857, 1:1000), and ZO-1 (abG041, 1:1000) were purchased from Abcam (Cambridge, UK). Images of tissue samples were acquired using an Olympus BX53 microscope (Tokyo, Japan). Image-Pro Plus v.6.0 software was used to analyze adipocyte diameters in the H&E-stained sections.
6
Immunohistochemical Staining of Immune Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue sections (5-μm thick) were placed on a slide and dried in the incubator at 37°C. After xylene dewaxing, the slices were immersed in 50%, 70%, and 80% ethanol for 2 min each and in eosin for 5 s. The specimens were then routinely dehydrated, made transparent, and fixed. The sections were washed with water, the antigen was heat repaired, the primary antibody was incubated, and the secondary IgG was incubated with appropriate biotin (1:1,000, #S0001, Affinity Biosciences, Wuhan, China). The slices were stained with hematoxylin for 5 min, rinsed with tap water for 3 min, rinsed with 1% hydrochloric acid for 2 s, rinsed with tap water for 2 min, and then sealed by gradient dehydration. For IF staining, the specimens were covered with a fluorescent-labeled antibody and stored in an enamel box for 30 min. Examination of co-stained sections and images were collected using a light microscope (Olympus, Tokyo, Japan). The corresponding primary antibodies were against CD80 (ab254579.html">ab254579 , Abcam, Cambridge, UK), CD86 (ab254579.html">ab119857 , Abcam), F4/80 (ab60343, Abcam), iNOS (ab178945, Abcam), CD4 (ab183685, Abcam), CD8 (ab237723, Abcam), CD3 (ab135372, Abcam), CD44 (ab243894, Abcam), and Il-12R (ab282729, Abcam).
7
Mapping M1 Macrophages in Pancreatic Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The distribution of the M1 phenotype in the pancreas was assayed by immunofluorescence staining. Pancreatic tissues were cut into sections which were dewaxed. CD68 (ab955, Abcam) expression was detected by anti‐mouse Alexa‐Flour647 (P0191, Biotechnology Co., Shanghai, China), and CD86 (ab119857, Abcam) expression was detected by anti‐rabbit FITC (Beyotime). Then, the samples were stained with DAPI to visualize the nuclei. The distribution of M1 macrophages in the pancreas was examined by a fluorescence microscope (Olympus Optical Co., Ltd, Tokyo, Japan).
8
Retinal Immunohistochemistry in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The eyes of mice at P17 were enucleated and fixed at RT for 1 h in 4% PFA. After removing the cornea and lens, which were fixed in 4% PFA overnight, the eyes were dehydrated in 20% sucrose and embedded in OCT compound. Then, they were frozen at −20 °C overnight and cut into 10 μm thick sections. Next, the sections were washed with PBS before permeabilizing and blocking with 1% BSA and 0.25% Triton-X-100 for 1 h at RT. The sections were incubated with IB4 (1:200), iba1 (1:500), GFAP (1:200), CD86 (1:100, ab119857, Abcam, Cambridge, UK), CD206 (1:100, AF2535, R&D systems, Minneapolis, MN, USA), and VEGF (1:100, NB100-664, Novus, Centennial, CO, USA) overnight at 4 °C. After washing in PBS, sections were incubated for 2 h at RT with a mixture of secondary antibodies and DAPI. The images of retinal frozen sections were also taken by confocal microscopy. Three fields of view (at ×400 magnification) on each section and at least three sections for each eye were observed. For statistical analysis of frozen sections, six eyes were assessed for each group.
9
Immunohistochemical analysis of tumor samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin-embedded samples were sectioned at 4-mm thickness. Antigen retrieval was performed using a pressure cooker for 3 min in 0.01 M citrate buffer (pH 6.0). Samples were blocked in PBS with 2% BSA for 1 h at room temperature and incubated with antibodies specific for Ki-67 (1:100; Abcam, ab16667), MMP9 (1:100; Abcam, ab283575), CD86 (1:100; Abcam, ab119857), CD206 (1:100; Abcam, ab64693), iNOS (1:100; Abcam, ab178945) and Arg1 (1:100; Abcam, ab96183) overnight at 4 °C. Alexa Fluor or HRP conjugated secondary antibodies was incubated for 1 h at room temperature. DAPI was then used for counterstaining the nuclei, and images were obtained by fluorescence microscope (Axio Observer D1, Zeiss).
10
Antibody Panel for Ferroptosis Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies composed of Rabbit anti‐human Glutathione Peroxidase 4 (GPX4, Abcam Cat#ab125066, RRID: AB_10973901), Rabbit anti‐human NOX1 (Abcam Cat# ab78016, RRID: AB_1566505), Rabbit anti‐human FACL4 (Abcam Cat# ab155282, RRID: AB_2714020), Rabbit anti‐CD4 (Abcam Cat# ab183685, RRID: AB_2686917), Rabbit anti‐CD8 (Abcam Cat# ab93278, RRID: AB_10563532), and Rabbit anti‐CD86 (Abcam Cat# ab119857, RRID: AB_10902800).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!