Sections (4 μm thick) from the paraffin-embedded colon tissue blocks were prepared for IHC or IF staining. Briefly, staining was performed with primary antibodies against TNF-α (1 : 600, rabbit polyclonal, Bioss, bs-2081R), IL-1β (1 : 2,000, rabbit polyclonal, ABclonal, A11369), NF-κB/p65 (1 : 5,000, rabbit polyclonal, Servicebio, GB11142), F4/80 (1 : 1000, CST, 70076), CD86 (1 : 500, Bioss, bs-1035R), and CD163 (1 : 300, Bioss, bs-2527R). Goat anti-rabbit IgG (HRP) (1 : 4,000, Abcam, ab205718), Alexa 488-conjugated donkey anti-rabbit IgG(H+L) (1 : 400, Life Technologies, A21206), and Cy3 (1 : 200) were used as secondary antibody staining reagents for IF staining, while a goat polyclonal secondary antibody recognizing mouse IgG (1 : 500, Abcam, ab150113) was used for immunohistochemistry; DAPI-containing TBST was applied for 5 min. Sections were visualized using an Olympus microscope (Olympus BX53, Olympus Corporation).
Bs 1035r
Manufactured by Bioss Antibodies
Sourced in United States
The Bs-1035R is a laboratory equipment product offered by Bioss Antibodies. It is designed for general laboratory use. The core function of the Bs-1035R is to perform essential tasks in a research or diagnostic setting. No further details about its intended use or capabilities can be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Bs 2081r, by Bioss Antibodies (2 mentions)
Ab150113, by Abcam (1 mentions)
Bs 2527r, by Bioss Antibodies (1 mentions)
A21206, by Thermo Fisher Scientific (1 mentions)
Ab205718, by Abcam (1 mentions)
Bs 2211r, by Bioss Antibodies (1 mentions)
Bs 0698r, by Bioss Antibodies (1 mentions)
Ab23990, by Abcam (1 mentions)
Ab157210, by Abcam (1 mentions)
Ab134189, by Abcam (1 mentions)
Ab9324, by Abcam (1 mentions)
Ab68672, by Abcam (1 mentions)
Ab25377, by Abcam (1 mentions)
Ab5103, by Abcam (1 mentions)
Nb100 39002, by Novus Biologicals (1 mentions)
Streptavidin apc, by BioLegend (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Immpact dab, by Vector Laboratories (1 mentions)
Bz x810, by Keyence (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
4 protocols using bs 1035r
1
Immunostaining of Colon Tissue
2
Comprehensive Protein Expression Analysis
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A total cell protein extraction kit (Milipore, Billerica, MA, USA) was used to extract total protein. An equivalent amount of protein from each sample was electrophoresed by 12% SDS-PAGE and transferred to nitrocellulose membrane. After being blocked, membranes were incubated with anti-TLR4 (1:1000; PA5-23124, Invitrogen), anti- MHC-I (1:1000; ab134189 and ab22367, Abcam), anti-MHC-II (1:1000; ab157210 and ab23990, Abcam), anti-CD80 (1:1000; PA5-19211, Invitrogen and bs-2211r, Bioss, Woburn, MA, USA), anti-CD86 (1:1000; bs-1035r, Bioss), anti-TNF-α (1:1000; bs-2081R, Bioss), anti-IL-6 (1:2000; ab9324, Abcam), anti-IL-10 (1:1000; bs-0698r, Bioss), anti-CXCL10 (1:1000; PAA371Ra01; Cloud-Clone Corp., Houston, TX, USA), anti-TGF-β1 (1:1000; c0340, Assay biotechnology, Sunnyvale, CA, USA) and TGF-β2 (1:1000; 5343r-100, BioVision, Milpitas, CA, USA) overnight at 4 °C. Membranes were then washed three times with PBS/0.1%Tween-20 (5 min each), and incubated with a corresponding secondary antibody (1:5000) for 2 h at room temperature. Bands were detected using a chemiluminescence ECL kit (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and were quantified using the Sigma-Gel software (Jandel Scientific Software, Sari Kafael, CA, USA).
3
Lung Tissue Histology and Immunostaining
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The mice were sacrificed and perfused intracardially with PBS and 4% paraformaldehyde. Then, the lung tissue was soaked in 4% formaldehyde for 24 h at 4 °C and then steeped in 30% sucrose solution until the tissue sank (about 48 h). The lungs were cut into 8-um-thick slices. The lung slides were stained with hematoxylin and eosin (H&E) and examined using light microscopy [25 (link)]. For immunofluorescent staining, the slides were preincubated in blocking solution. Then, the lung sections were incubated for over 12 h at 4 °C with the primary antibodies: anti-CitH3 (ab5103, Abcam, Cambridge, UK), anti-Ly6g (ab25377, Abcam), anti-NE (ab68672, Abcam), CD16 (PA5-47230, Invitrogen, Waltham, MA, USA) and CD86 (BS-1035R, Bioss, Woburn, MA, USA). Then, the lung sections were incubated with secondary antibodies at room temperature for 2 h. At last, the sections were covered with DAPI and observed using a fluorescence microscope. At least three sections per mice were examined.
4
Immunofluorescence Analysis of Immune Markers
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The frozen sections were rinsed with Tris-buffered saline (TBS) at room temperature for 10 min. After blocking with 10% normal goat serum (NGS) containing blocking solution (10% NGS/TBS) for 1 h, each antibody was reacted in the humid chamber at 4 °C overnight. The concentration of each antibody used in the experiments was 1:500 for CD86 (bs-1035R: Bioss, Boston, MA, USA), 1:1000 for Foxp3 (NB100-39002: Novus Biologicals, Centennial, CO, USA) and 1:500 for Axin2 (ab32197: Abcam, Cambridge, UK). After the reaction with these primary antibodies, the bound antibodies were visualized with a VECTASTAIN Elite ABC kit (VECTOR LABORATORIES, Burlingame, CA, USA) and Diaminobenzidine (DAB) (ImmPACT DAB: VECTOR LABORATORIES). The tissues were counter stained with Mayer’s hematoxylin solution, and then dehydrated, penetrated, and sealed with MOUNT-QUICK.
For immunofluorescence analysis, the sections were sequentially reacted with the anti-CD68 antibody (1:500, ab125212: Abcam) and then reacted with the biotinylated secondary antibody. After the reactions, the sections were reacted with APC-streptavidin (405,207: BioLegend, San Diego, CA, USA) and the FITC-anti-CD206 antibody (1:200: bs-4747R-FITC, Bioss). Finally, the sections were stained with DAPI (Sigma) and observed under a fluorescence microscope (BZ-X810: KEYENCE).
For immunofluorescence analysis, the sections were sequentially reacted with the anti-CD68 antibody (1:500, ab125212: Abcam) and then reacted with the biotinylated secondary antibody. After the reactions, the sections were reacted with APC-streptavidin (405,207: BioLegend, San Diego, CA, USA) and the FITC-anti-CD206 antibody (1:200: bs-4747R-FITC, Bioss). Finally, the sections were stained with DAPI (Sigma) and observed under a fluorescence microscope (BZ-X810: KEYENCE).
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