In kidney tissues and HK-2 cells, RIPA lysis buffer (20–188, Sigma-Aldrich) was used for extracting proteins. An equal quantity of protein was electrophoresed using SDS-PAGE and then transferred onto PVDF membranes. At 4 °C, primary antibodies against ELF4, Nrf2, NQO-1, GRP78, KIM-1, and IL-6 (1:1000, ab96075, ab137550, ab34173, ab21685, ab302932, and ab259341, Abcam, USA); IL-1β (1:1000, sc-12742, Santa Cruz Biotechnology); IL-18 and N-GSDMD (1:1000, A1115 and A22523, ABclonal, USA); Bax, Bcl-2, TNF-α, caspase-4, caspase-11, Caspase-12, CHOP, HO-1, and GSDMD (1:1000, 2772, 3498, 3707, 4450, 14340, 35965, 2895, 43966, and 39754, Cell signaling Technology, USA), and GAPDH (1:5000, 5174, Cell signaling Technology, USA) were deal with membranes overnight. After that, secondary antibodies (1:50000, 7074, Cell signaling Technology) were handled with membranes for 1 h. At last, protein bands were visualized by way of an enhanced chemiluminescence detection kit (Beyotime Biotechnology, China). Image J software (USA) was used to quantify protein expression.
A1115
Manufactured by ABclonal
Sourced in United States, China
The A1115 is a lab equipment product. It is designed for performing specific laboratory functions. No further details are available.
Automatically generated - may contain errors
Lab products found in correlation
A0964, by ABclonal (2 mentions)
Ab34173, by Abcam (1 mentions)
Ab259341, by Abcam (1 mentions)
Enhanced chemiluminescence detection kit, by Beyotime (1 mentions)
Ab21685, by Abcam (1 mentions)
Ripa lysis buffer, by Merck Group (1 mentions)
Sc 12742, by Santa Cruz Biotechnology (1 mentions)
A22523, by ABclonal (1 mentions)
Ab137550, by Abcam (1 mentions)
A5258, by ABclonal (1 mentions)
A0980, by ABclonal (1 mentions)
A16288, by ABclonal (1 mentions)
A10240, by ABclonal (1 mentions)
Protein extraction kit, by Beyotime (1 mentions)
Fibronectin, by ABclonal (1 mentions)
A5786, by ABclonal (1 mentions)
A12694, by ABclonal (1 mentions)
Ac026, by ABclonal (1 mentions)
A16678, by ABclonal (1 mentions)
Ap0727, by ABclonal (1 mentions)
6 protocols using a1115
1
Protein Expression Analysis in Kidney Tissues
2
Western Blot Analysis of Gastro Tissue Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples were extracted from 0.5 g of gastro tissue using a protein extraction kit (Beyotime Biotechnology, China); these samples were then used for western blot analysis, which was performed according to the manufacturer’s instructions. A dilution ratio of 1:1000 was used for all the primary antibodies, including rabbit anti-NLPP3 polyclonal antibody (19771–1-AP, Proteintech), rabbit anti-TLR4 polyclonal antibody (A5258, Abclonal), rabbit anti-p65 monoclonal antibody (CST 8242T, CST, Danvers, MA, USA), rabbit anti-GSDMD monoclonal antibody (CST 69469S, CST), rabbit anti-Caspase 1 monoclonal antibody (CST 3866T, CST, Danvers), mouse anti-iκBα monoclonal antibody (CST 4814T, CST), rabbit anti-Myd88 polyclonal antibody (A0980, Abclonal), rabbit anti-IL-1β polyclonal antibody (A16288, Abclonal), rabbit anti-IL-18 polyclonal antibody (A1115, Abclonal), rabbit anti-ZO-1 monoclonal antibody (CST 13,663, CST), rabbit anti-Occludin monoclonal antibody (CST 91,131, CST), rabbit anti-RORγ polyclonal antibody (A10240 Abclonal), rabbit anti-Foxp3 monoclonal antibody (CST12632; CST), and mouse anti-β-actin monoclonal antibody (CST4970; CST). A dilution ratio of 1:2000 was used for all the secondary antibodies, including anti-rabbit IgG, HRP-linked antibody (CST7074, CST). Enhanced chemiluminescence was used for detection.
3
Immunoblot Assay of Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblot assays were performed according to standard protocol. Target proteins were detected with antibodies against NLRP3 (A12694, Abclonal, Wuhan, China), collagen I (A5786, Abclonal, Wuhan, China), fibronectin (A16678, Abclonal, Wuhan, China), Flag (F1804, Sigma, St Louis, USA), caspase1 (A0964, Abclonal, Wuhan, China), IL-1β (A11369, Abclonal, Wuhan, China), IL-18 (A1115, Abclonal, Wuhan, China), phos-smad3 (AP0727, Abclonal, Wuhan, China), E-cadherin (3195, CST, Danvers, USA), N-cadherin (14215, CST, Danvers, USA), FIP1 (A7138, Abclonal, Wuhan, China), β-actin (AC026, Abclonal, Wuhan, China).
4
Western Blot Analysis of Inflammasome Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lysates were separated by SDS-PAGE before being transferred to a polyvinylidene difluoride (PVDF) membrane (ISEQ00010, Sigma-Aldrich, USA), which was then blocked for 1 hour at room temperature using blocking solution (5% skim milk and 0.05% Tween 20 in TBST). The membrane was incubated overnight at 4℃ with primary antibodies to caspase-1 (A0964, Abclonal), IL-1β (A11370, Abclonal), IL-18 (A1115, Abclonal), NLRP3 (A14223, Abclonal), NF-κB P65 (AF5006, Affinity), p-NF-κB p65 (AP0124, Abclonal) at dilutions of 1:2000 and rabbit anti-β-actin (AP0124, Abclonal) monoclonal antibodies at dilution of 1:50000, respectively. The membrane was then incubated for 1 h with the secondary antibody (goat anti-rabbit immunoglobulin (IgG) (H+L; s0001, acquired from Affinity) at a dilution of 1:5000.
After three washes with TBST, the signals were developed using the Hypersensitive ECL Western HRP Substrate kit (17046, Zen-bio).
After three washes with TBST, the signals were developed using the Hypersensitive ECL Western HRP Substrate kit (17046, Zen-bio).
5
Quantification of Hippocampal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue lysates were extracted from mouse hippocampus samples with lysis buffer (70-WB020, LiankeBio, Hangzhou, China) containing protease inhibitors. After quantification using a bicinchoninic acid kit (70-PQ0012, LiankeBio, Hangzhou, China), 30 μg of total protein was separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis at 75–120 V for 90 min and transferred to a polyvinylidene fluoride membrane at 300 mA for 30 min. After being blocked in 5% bovine serum albumin (BSA) for 1 h, the membrane was incubated overnight at 4 °C with primary antibodies against NLRP3 (1:1 000, ab270449, Abcam), IL-1β (1:1 000, ab9722, Abcam), IL-18 (1:1 000, A1115, Abclonal), amyloid-β1-42 (Aβ1-42) (1:1 000, YT5773, Immunoway), and β-actin (1:5 000, AF7018, Affinity). The membrane was then washed three times for 10 min each and incubated in the secondary antibody for 2 h at room temperature. Protein bands were visualized using an imaging system (ChemiScope 6100, ClinX, China). The integrated gray values of each band were measured using ImageJ (National Institutes of Health, Bethesda, MD, USA) to determine the relative protein expressions.
6
Western Blot Analysis of NLRP3 Inflammasome Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples were extracted using lysis buffer containing 1% protease inhibitor (Roche, Switzerland). Protein concentrations were measured by a BCA protein kit (Beyotime Institute of Biotechnology, Shanghai, China). The proteins were separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes (PALL, USA). The membranes were incubated with the antibodies against NLRP3 (1:1000; #ab97051, Abcam, Cambridge, UK), GSDMD (1:500; #ab209845, Abcam), IL-1β (1:500; #ab9722), caspase-1 (1:500; #WL02996a, Wanleibio, China), IL-18 (1:500; #A1115, Abclonal, Wuhan, China), and β-actin (1:1000; #bs-0061R, BIOSS, Boston, MA, USA) at 4 °C overnight and then with a fluorescence-conjugated anti-rabbit IgG secondary antibody (1:10,000; LI-COR Bioscience, Lincoln, NE, USA) for 50 min in the dark. The gray value of each protein was quantified by an Infrared Fluorescence Imaging Detector (LI-COR Bioscience) and normalized to β-actin as an internal control.
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!