Fucoidan Fuc1 (Undaria pinnatifida source, batch# 572001015) was obtained from Qingdao bright moon seaweed group co., LTD. (Qingdao, China). Samples Fuc2 (Undaria pinnatifida source, Lot# SLCK7680) and Fuc3 (Fucus Vesiculosis source, Lot# SLCJ3576) were purchased from Sigma-Aldrich (MO, USA). Murine RAW 264.7 cells were obtained from Biofavor Biotech (Wuhan, China). High glucose Dulbecco’s modified Eagle’s medium (DMEM) (with 4500 mg/L d -glucose, 3700 mg/L sodium bicarbonate, 584 mg/L l -glutamine, 110 mg/L sodium pyruvate and 15 mg/L phenol red), Fetal bovine serum (FBS), streptomycin and penicillin Trizol reagent were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Cytoplasmic and nuclear protein extraction kit, and nitric oxide (NO) detecting kit were obtained from Beyotime (Haimen, China). cDNA Synthesis SuperMix TransScript® and One-Step gDNA Removal were purchased from Transgen Biotechnology Co. (Beijing, China). Fluorescence real-time quantitative PCR premix (SYBR) and total RNA isolation kit were purchased from Tiangen Biotechnology Co. (Beijing, China). GAPDH, iNOS, COX-2 antibodies and HRP-linked anti-rabbit IgG (secondary antibody) were purchased from Abcam (Shanghai, China). The monosaccharide standards were obtained from BoRui Saccharide Biotech Co. Ltd (Yangzhou, China). Other chemical reagents were of analytical grade.
Cox 2 antibody
Manufactured by Abcam
Sourced in United Kingdom, China
The COX-2 antibody is a laboratory reagent used for the detection and quantification of the cyclooxygenase-2 (COX-2) enzyme in biological samples. COX-2 is an important enzyme involved in the inflammatory response and is a common target for research in various fields, such as immunology, oncology, and neuroscience.
Automatically generated - may contain errors
Lab products found in correlation
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Gapdh, by Abcam (1 mentions)
Total rna isolation kit, by Tiangen Biotech (1 mentions)
T aoc test kit, by Nanjing Jiancheng (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Hrp coupled secondary antibodies, by Promega (1 mentions)
Dmso d6, by Merck Group (1 mentions)
Ecl kit, by Abcam (1 mentions)
Trypsin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Cdcl3, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Pvdf membrane, by Pall Corporation (1 mentions)
P stat1, by Santa Cruz Biotechnology (1 mentions)
P stat3, by Santa Cruz Biotechnology (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
N cadherin antibody, by Cell Signaling Technology (1 mentions)
E cadherin antibody, by Cell Signaling Technology (1 mentions)
Gapdh mouse monoclonal antibody, by ABclonal (1 mentions)
Recombinant human il 1β, by Sangon (1 mentions)
13 protocols using cox 2 antibody
1
Fucoidan Extraction and Characterization
2
Intestinal Barrier Markers Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Diamine oxidase activity (DAO) detection kit was purchased from Beijing Solarbio Technology Co., Ltd. A total antioxidant capacity (T-AOC) test kit was obtained from Nanjing Jiancheng Biological Engineering Research Institute Co., Ltd. ELISA kits (Shanghai Langton Biotechnology Co., Ltd.)were used to measure D-lactate, a mechanical barrier marker, and SIgA, a mucosal barrier marker. Claudin-1, proliferating cell nuclear antigen (PCNA), TNF-α and GAPDH antibodies were provided by Wuhan Proteintech Biotechnology Co., Ltd. Occludin, NF-κB (P65) and COX-2 antibodies were purchased from Abcam (Cambridge Science Park in Cambridge, UK). Mouse IL-10 and IL-1β ELISA kits was purchased from Beijing Solarbio Technology Co., Ltd.
3
Optimized Western Blot Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s modified Eagle’s medium (DMEM) and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, Missouri, USA). PVDF membranes were purchased from Pall Corporation (Ann Arbor, MI, USA). Actin and Cox-2 antibodies were purchased from Abcam (Cambridge, MA, USA). HRP-coupled secondary antibodies were purchased from Promega Corporation (Madison, WI, USA). The ECL kit was purchased from Abcam (Milton, Cambridge, UK). Fetal bovine serum (FBS), penicillin-streptomycin, glycine, lysis buffer solution, phosphate buffer saline (PBS), bovine serum albumin (BSA), tris-buffered saline with Tween 20® (TBST), HEPES buffer, sodium dodecyl sulfate (SDS), well plates, pentane, diethyl ether, hexane, ethyl acetate, anisaldehyde, ethanol, paraformaldehyde, trypan blue, Ficoll (Histopaque-1077). Crystal violet, trypsin, d6-DMSO and CDCl3 were purchased from Sigma (St. Louis, Missouri, USA) unless otherwise stated.
4
TLR-4, COX-2, and STAT Signaling in RAW 264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression of TLR-4 and COX-2 in the RAW 264.7 cells were analyzed using flow cytometry. The pSTAT1 and pSTAT3 were also analyzed to explore the related signalling pathway by which the extracts acted on the cells.
After treatment, RAW 264.7 cells in each group were dissociated to pellets (1 × 105 each) and washed twice with PBS, and then the TLR-4 antibody (eBioscience, Shanghai, China) was added to the cells. After staining for 10 min, the pellets were analyzed by flow cytometry. By using fixation/permeabilization concentrate (Affymetrix eBioscience, San Diego, CA) pre-permeated, the remaining prepared pellets were stained with pSTAT1, pSTAT3 (Santa Cruz Biotechnology, Dallas, TX) or COX-2 antibodies (Abcam, Shanghai, China) for 10 min to test the pSTATs and COX-2 levels.
After treatment, RAW 264.7 cells in each group were dissociated to pellets (1 × 105 each) and washed twice with PBS, and then the TLR-4 antibody (eBioscience, Shanghai, China) was added to the cells. After staining for 10 min, the pellets were analyzed by flow cytometry. By using fixation/permeabilization concentrate (Affymetrix eBioscience, San Diego, CA) pre-permeated, the remaining prepared pellets were stained with pSTAT1, pSTAT3 (Santa Cruz Biotechnology, Dallas, TX) or COX-2 antibodies (Abcam, Shanghai, China) for 10 min to test the pSTATs and COX-2 levels.
5
Cardiac Biomarker Evaluation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
Acorus tatarinowii, propranolol hydrochloride tablets, and isoproterenol hydrochloride injection were purchased from Hebei Jinye Pharmaceutical Co. Ltd. (Hebei, China), Jiangsu Yabang Epson Pharmaceutical Co. Ltd. (Jiangsu, China), and Shanghai Wellhope Pharmaceutical Co. Ltd. (Shanghai, China), respectively. SOD, LDH, COX-2, and PPARα ELISA kits were obtained from Jiangsu Meimian Industry Co., Ltd. (Jiangsu, China), while COX-2 antibodies and GADPH antibodies were purchased from Abcam (Cambridge, England). PPAR-α antibodies were bought from Beijing Boaosen Biotechnology Co. Ltd. (Beijing, China).
+ Expand
Acorus tatarinowii, propranolol hydrochloride tablets, and isoproterenol hydrochloride injection were purchased from Hebei Jinye Pharmaceutical Co. Ltd. (Hebei, China), Jiangsu Yabang Epson Pharmaceutical Co. Ltd. (Jiangsu, China), and Shanghai Wellhope Pharmaceutical Co. Ltd. (Shanghai, China), respectively. SOD, LDH, COX-2, and PPARα ELISA kits were obtained from Jiangsu Meimian Industry Co., Ltd. (Jiangsu, China), while COX-2 antibodies and GADPH antibodies were purchased from Abcam (Cambridge, England). PPAR-α antibodies were bought from Beijing Boaosen Biotechnology Co. Ltd. (Beijing, China).
6
Reagents and Antibodies for Cell Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DMEM/F12 medium (#SH30023.01) was from Hyclone (Shanghai, China) and fetal bovine serum (FBS; #04–001-1A) was bought from Biological Industries (BI; Cromwell, CT, USA). 17β-estradiol/estrogen/E2 (#E2758) and lipopolysaccharides (LPS; #L4391) were from Sigma-Aldrich (Shanghai, China). Recombinant Human IL-1β (#C600002) was bought from Sangon Biotech (Shanghai, China). GAPDH mouse monoclonal antibody (#40493) was procured from ABclonal (Boston, USA). Cytokeratin 18 antibodies (#4548), E-cadherin antibodies (#14472), Vimentin antibodies (#5741), N-cadherin antibodies (#13116), Progesterone Receptor (PR) antibodies (#8757S), and ERα antibodies (#13258) were acquired from Cell Signaling Technology (Danvers, MA, USA). HSD17B1 antibodies (#ab51045), TERT antibodies (#ab32020), ERβ antibodies (#ab288), COX-2 antibodies (#ab15191) and CYP17A1 antibodies (#ab125022) were purchased from Abcam (Cambridge, UK).
7
Maintenance and Characterization of TNBC Cell Line MDA-MB-231
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human triple-negative breast cancer (TNBC) cells MDA-MB-231 were maintained in DMEM (Hyclone, Cramlington, UK) supplemented with antibiotics (penicillin 50 U/mL; streptomycin 50 µg/mL) and 10% fetal bovine serum (Hyclone, Cramlington, UK) at 37 °C. The culture medium was changed every three days, and cells were passaged once a week when the culture reached 95% confluence. Cell viability measured using trypan blue dye exclusion was higher than 99% in all experiments. All in vitro experiments were repeated at least three times. Antibodies to PD-L1, Akt, phospho-Akt, phospho-ERK, Snail, p21, and β-tubulin were obtained from Cell Signaling Technology (Cell Signaling, Beverly, MA, USA). Antibodies to ERK, Survivin, RhoA, Rac1, Cdc42, c-Fos, c-Myc, and β-actin HRP were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). COX2 antibody was obtained from Abcam (Cambridge, UK).
8
Immunohistochemical Detection of COX-2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lungs embedded in paraffin and sectioned, and the sections treated with COX-2 antibody (Abcam, Cambridge, UK) overnight. The slides were then treated with secondary antibodies and incubated with DAB substrate to detect COX-2 expression, as described previously [26 (link)].
9
Intestinal Protein Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total Protein Extraction Kit (KeyGEN Bio TECH, Nanjing, China) was used for total protein isolation from full thickness of intestinal wall. BCA Protein Assay Kit (Beyotime Institute of Biotechnology, Haimen, China) was used for protein concentration determination. The blots on PVDF membrane were respectively probed with corresponding protein antibodies including MLCK antibody [1:1000; Abcam (Hong Kong) Ltd.], claudin-2 antibody (1:800; Santa Cruz Biotechnology Co. Ltd., Shanghai, China), iNOS antibody [1:1000; Abcam (Hong Kong) Ltd.], COX-2 antibody [1:1000; Abcam (Hong Kong) Ltd.], and NF-KappaB-p65 antibody [1:1000; Abcam (Hong Kong) Ltd.], respectively. Multi Spectral imaging system (UVP, Cambridge, UK) was used to detect and quantify the bands for special protein determination.
10
Lung Histopathology and COX-2 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were sacrificed at 0, 3, 7, 15, and 30 days after infection with MS strains and lung tissue was collected from individual mice. Lung tissues were fixed in 10% formalin and then dehydrated, paraffin-embedded, and sectioned at 5µm. Sections of lung tissues were stained with haematoxylin and eosin for histopathological analysis. Morphometric image analysis was performed at ×100 or ×200 magnification. For immunohistochemical staining of COX-2, paraffin-embedded lung sections were stained using rabbit anti-mouse polyclonal COX-2 antibody (Abcam, Cambridge, UK), biotin-conjugated rabbit anti-mouse IgG, and a peroxidase-conjugated streptavidin antibody. Then, visualization was performed with 3,3-diaminobenzidine as the peroxidase substrate (Sigma-Aldrich).
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!