The following reagents were purchased from Beijing Bioss Biotechnology: c-fos rabbit anti-mouse polyclonal antibody, ZO-1 rabbit anti-mouse polyclonal antibody, Occludin rabbit anti-mouse polyclonal antibody, myosin light chain kinase (MLCK) rabbit anti-mouse polyclonal antibody, P-glycoprotein (P-gp) rabbit anti-mouse polyclonal antibody, multidrug resistance associated protein 1 (MRP1) rabbit anti-mouse polyclonal antibody, multidrug resistance associated protein 2 (MRP2) rabbit anti-mouse polyclonal antibody, goat anti-rabbit immunohistochemistry kit, and hematoxylin. Other reagents used were as follows: glacial acetic acid, analytical grade (Beijing Chemical Works, Batch No. 20100603); osmic acid, acetone, embedding media, uranyl acetate stain, and lead citrate stain (supplied by the Experimental Research Center); potassium permanganate, analytical grade (Beijing Chemical Works, Batch No. 820308); toluidine blue, analytical grade (Sinopharm Chemical Reagents, Batch No. WC20050120); anhydrous ethanol, analytical grade (Beijing Chemical Works, Batch No. 20110); xylene, analytical grade (Sinopharm Chemical Reagents, Batch No. 20106); neutral gum (Sinopharm Chemical Reagents, Batch No. 20120320); hematoxylin (Beyotime Biotechnology, Batch No. C0105); ethylenediamine tetraacetic acid (EDTA), analytical grade (Beijing Chemical Works, Batch No. 840529).
Neutral gum
Manufactured by Sinopharm
Sourced in China
Neutral gum is a lab equipment product that serves as an adhesive, binding agent, and emulsifier. It is a versatile and inert substance used in various scientific and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Xylene, by Sinopharm (5 mentions)
Toluidine blue, by Sinopharm (3 mentions)
Paraffin, by Sinopharm (2 mentions)
Glacial acetic acid, by Sinopharm (2 mentions)
Anhydrous ethanol, by Beijing Chemical Works (1 mentions)
Glacial acetic acid, by Beijing Chemical Works (1 mentions)
Hematoxylin, by Beyotime (1 mentions)
Ethylenediamine tetraacetic acid edta, by Beijing Chemical Works (1 mentions)
Glycerin, by Sinopharm (1 mentions)
Hematoxylin, by Sinopharm (1 mentions)
Ethyl alcohol, by Sinopharm (1 mentions)
Chloral hydrate, by Sinopharm (1 mentions)
Citric acid, by Sinopharm (1 mentions)
Eclipse e100, by Nikon (1 mentions)
Mayer hematoxylin, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Triton x 100, by Keygen Biotech (1 mentions)
Goat anti rabbit igg hrp, by Keygen Biotech (1 mentions)
Methanol, by Sinopharm (1 mentions)
Ethanol, by Sinopharm (1 mentions)
12 protocols using neutral gum
1
Immunohistochemical Analysis of Cellular Markers
2
Preparation and Characterization of Plant Soot
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Plant soot (Lot#2-9243-2-01-11; purity 95%) was provided by China National Center for Veterinary Drug Safety Assessment (Beijing, China). 2,4,6-Trinitrobenzene (≥99% pure, CAS: 88-89-1) was obtained from Beijing Giant-Carrier Co., Ltd. (Beijing, China); 37–40% formalin and carboxyl methyl cellulose sodium (CMC-Na) were purchased from Tianjin Damao Chemical Reagent Factory Co., Ltd. (Tianjin, China). Potassium hydroxide was purchased from Shantou Xilong Chemical Plant Co., Ltd. (Shantou, China). Ethyl alcohol, xylene, paraffin, neutral gum, transparent agent, hematoxylin, eosin, chloral hydrate, citric acid, chromium potassium sulfate, glacial acetic acid, glycerin, and chloral hydrate were obtained from Sinopharm Chemical Reagent Co., Ltd. (Beijing, China). Alizarin red and sodium hydrochloride were purchased from Beijing Chemical Reagent Co., Ltd. (Beijing, China). Serum albumin (Alb), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN), total cholesterol (TCH), creatinine (Cr), glucose (Glu), total protein (TP), and triglyceride (TG) detection kits were all obtained from Shanghai Kehua Bio-Engineering Co., Ltd. (Shanghai, China).
3
Quantifying Spinal Cord Motor Neurons
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We used Nissl staining to identify spinal cord motor neurons. A decrease in the number of Nissl bodies and abnormal motor neuron morphology indicated that nerve cells may be damaged. Paraffin sections of the lumbosacral enlargements of the spinal cord were deparaffinized and rehydrated, and the sections were stained with toluidine blue (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) for 5 minutes, washed with water, and differentiated with 1% glacial acetic acid (Sinopharm Chemical Reagent Co., Ltd.). The reaction was terminated with tap water. The degree of stain differentiation was controlled by observation under an upright microscope (ECLIPSE E100, Nikon). After washing with water, the sections were dried in an oven (Sinopharm Chemical Reagent Co. Ltd.), cleared with xylene for 5 minutes, and mounted with neutral gum (Sinopharm Chemical Reagent Co., Ltd.). Serial coronal slides of the paraffin-embedded lumbosacral enlargements of the spinal cord were obtained and analyzed at intervals of six sections. The section thickness was 5 μm. The slides were scanned using an upright microscope (ECLIPSE E100, Nikon) with a 40× objective lens. The number of motor neurons in the lateral nucleus of the anterior horn of the spinal cord was manually calculated in three non-overlapping areas (220 × 350 μm2).
4
Histological Staining of Cell Samples
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The cells were fixed with 4% paraformaldehyde for 15 minutes, and Mayer hematoxylin (H9627; Sigma, Shanghai, China) was added dropwise and stained on the coverslip for 5 minutes with 1% water-soluble eosin (71014544; Sinopharm Group, Beijing, China). After staining for 2 to 4 minutes, the slides were mounted with neutral gum (10004160; Sinopharm Group) after treatment with absolute ethanol (10009218; Sinopharm GroupChina) and xylene (10023418; Sinopharm Group).
5
Investigating LPS-Induced Inflammation in Mice
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Male C57BL/6 mice with average weights of 23–26 g were purchased from the Corporation of Lingchang Biological Technology (Shanghai, China). The animals were allowed to acclimate to their surroundings for 1 week. LPS was purchased from Sigma (USA). LFM-A13 was purchased from Topscience (Shanghai, China). Kits for protein extraction, BCA protein assay, SDS-PAGE gel preparation, Western blot analysis, hematoxylin and eosin (H&E staining), TUNEL apoptosis assay, mouse IL-4 enzyme-linked immunosorbent assay (ELISA), mouse IL-6 ELISA, mouse TNF-α ELISA, and myeloperoxidase (MPO) colorimetric activity assay were purchased from KeyGEN (Nanjing, China). Neutral gum, methanol, ethanol, and xylene were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Triton-X100 was purchased from KeyGEN (Nanjing, China). Rabbit anti-GAPDH (10B8) and goat antirabbit IgG-HRP were purchased from KeyGEN (Nanjing, China). Rabbit anti-p-BTK (ab52192) was purchased from Abcam (Shanghai, China). Rabbit anti-BTK (DF6472) was purchased from Affinity Biosciences (Cincinnati, USA).
6
Immunohistochemical Staining Protocol
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The EliVision Plus kit and 3,3′-diaminobenzidine (DAB) kit were obtained from Fuzhou Maixin Biotechnical Co., Ltd. (Fuzhou, China). DAB-1031 and hematoxylin staining solution were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Neutral gum was obtained from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Polylysine solution, phosphate-buffered saline (PBS) and sodium citrate buffer were obtained from Nanjing KeyGen Biotech. Co., Ltd. (Nanjing, China).
7
Histological Analysis of Fixed Tissue
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Fresh tissue was fixed in 4% paraformaldehyde for over 24 h. After dehydration and paraffin embedding, the tissue was cut to a thickness of 4 μm. After paraffin sections were dewaxed, H&E staining was presented. The sections were stained with hematoxylin (G1004, Servicebio, Wuhan, China) for 5 min and 0.5% eosin solution (G1001, Servicebio) for 2 min. For Masson's staining, the sections were soaked in iron hematoxylin staining solution (G1006-2/3, Servicebio) for 3 min and Lichun acid magenta staining solution (G1006-4, Servicebio) for 5 min. After rinsing with tap water, the sections were stained with phosphomolybdic acid (G1006-5, Servicebio) for 3 min, aniline blue (G1006-6, Servicebio) for 5 min, and differentiated with 1% glacial acetic acid for 1 min. Following dehydration, the sections were mounted with neutral gum (10004160, Sinopharm, Shanghai, China) and placed under an optimal microscope (Nikon Eclipse CI, Nikon, Japan) for observation.
8
Histological Analysis of Bone Tissue
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The bone tissue was decalcified with 10% EDTA (Solarbio), dehydrated with an alcohol gradient (Sinopharm, China), made transparent using xylene (Sinopharm), and embedded in paraffin (Sinopharm). The bone tissue was sliced with a Leica pathological slicer (RM2016, Leica, Germany) and baked at 60 °C for 3 h. The slices were dewaxed in xylene and gradient alcohol in turn. The dewaxed sections were stained with Mayer’s hematoxylin and 1% water-soluble eosin solution (Solarbio). The slices were dehydrated in graded alcohol, rendered transparent in xylene, and finally sealed with neutral gum (Sinopharm). Images were collected by using a BX53 microscope (Olympus, Japan).
9
Eriochrome Cyanine Staining Protocol
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Slides were washed in 1X PBS three times, and dyed with eriochrome cyanine (catalog no. B100835-5G, Shanghai Aladdin Bio-Chem Technology Co., Ltd., Shanghai, China) for 30 min, and then washed three times with 1X PBS. Sections were differentiated by 10% iron alum (catalog no. 10009218, Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) for 8 min, and washed three times with 1X PBS, prior to coverslipping with neutral gum (catalog no. 1000416, Sinopharm Chemical Reagent Co., Ltd.).
10
Histopathological Analysis of Mouse Lungs
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Mouse lung tissue was first soaked and fixed with 4% paraformaldehyde for 1 week, embedded in xylene (Cat#10023418, Shanghai, China, Sinopharm Group Chemical Reagent Co., Ltd.), stained with hematoxylin (Cat#517-28-2, MACKLIN) for 5 min, stained with eosin (Cat#15086-94-9, MACKLIN) for 3 min, and then the tissue sections were sealed in gradient ethanol (Cat#100092683, Sinopharm Group Chemical Reagent Co., Ltd.) and neutral gum (Cat#10004160, Sinopharm Group Chemical Reagent Co., Ltd.) to observe the histopathological changes of the mouse lung tissue under the microscope.
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