At 0, 4, 8, 12, 24, and 48 h post-infection, protein lysate samples were boiled for 10 min in sodium dodecyl sulfate (SDS) buffer. The samples were then separated on a 12% SDS-polyacrylamide gel electrophoresis gel and electro-transferred to a nitrocellulose membrane (Millipore, USA) for 40 min. The membrane was blocked with 5% non-fat milk (Thermo Scientific, USA) for 1 h, and then incubated with rabbit anti-SUMO1 or Ubc9 monoclonal antibodies (Abcam, UK) diluted in Tris-buffered saline with Tween 20 (TBST) buffer for 1 h at 37°C. The membrane was washed with TBST buffer 3 times and incubated with HRP-conjugated mouse anti-rabbit IgG (Bioworld, USA) overnight at 4°C. After 3 washes, the membrane was stained using an HRP-DAB kit (Zhongshan Golden Bridge, China).
Hrp dab kit
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
The HRP-DAB kit is a laboratory product for the detection and visualization of target proteins in biological samples. It utilizes the horseradish peroxidase (HRP) enzyme and 3,3'-diaminobenzidine (DAB) chromogen to facilitate the localization and identification of specific proteins through immunohistochemical or immunocytochemical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated mouse anti rabbit igg, by Bioworld Technology (2 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Non fat milk, by Thermo Fisher Scientific (1 mentions)
Rabbit monoclonal anti caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti mouse igg, by Bioworld Technology (1 mentions)
Hrp conjugated goat anti rabbit igg, by Bioworld Technology (1 mentions)
Monoclonal rabbit anti nlrp3 antibody, by Cell Signaling Technology (1 mentions)
Evo m mlv reverse transcription kit, by Accurate Biology (1 mentions)
Immunohistochemical staining kit, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Ag rnaex pro rna kit, by Accurate Biology (1 mentions)
Sybr green pro taq hs kit, by Accurate Biology (1 mentions)
4 protocols using hrp dab kit
1
SUMO Protein Expression Dynamics
2
Western Blot Analysis of Autophagy and Inflammasome Markers
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Protein lysate samples were separated by 12% SDS-PAGE gel and transferred to the NC membrane for 40 min (100 mM of Tris-HCl, 150 Mm of NaCl, 0.05% Tween 20, and pH 7.2). The NC membrane was blotted with 5% non-fat milk in TBST for 1 h, followed by incubation with a primary antibody diluted with TBST for 2 h at 37°C. The membrane was washed with TBST 3 times and incubated with a secondary antibody at 37°C for 2 h. After washing with PBS 3 times, the NC membrane was stained using a HRP-DAB kit (Zhongshan Golden Bridge, Beijing, China).
The primary antibodies that we used are: mouse anti-p62 monoclonal antibody (Abcam, UK), rabbit anti-LC3A/B monoclonal antibody, rabbit anti-NLRP3 monoclonal antibody, rabbit anti-Caspase-3 monoclonal antibody (Cell Signaling Technology, USA), rabbit anti-caspase-1 p10 monoclonal antibody, and rabbit anti-β-actin monoclonal antibody (Jackson ImmunoResearch, USA). Secondary antibodies were Cy3-conjugated donkey anti-rabbit monoclonal antibody (Jackson ImmunoResearch, USA), HRP-conjugated goat anti-rabbit IgG, and HRP-conjugated goat anti-mouse IgG (Bioworld, USA).
The primary antibodies that we used are: mouse anti-p62 monoclonal antibody (Abcam, UK), rabbit anti-LC3A/B monoclonal antibody, rabbit anti-NLRP3 monoclonal antibody, rabbit anti-Caspase-3 monoclonal antibody (Cell Signaling Technology, USA), rabbit anti-caspase-1 p10 monoclonal antibody, and rabbit anti-β-actin monoclonal antibody (Jackson ImmunoResearch, USA). Secondary antibodies were Cy3-conjugated donkey anti-rabbit monoclonal antibody (Jackson ImmunoResearch, USA), HRP-conjugated goat anti-rabbit IgG, and HRP-conjugated goat anti-mouse IgG (Bioworld, USA).
3
Brucella Infection Protein Analysis
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According to the Brucella infection models, at 0, 2, 4, 8, 12 and 24 h after infection, protein lysate samples were separated by 12% SDS-PAGE gel and transferred to the Nitrocellulose membrane (NC) membrane for 40 min (100 mM of Tris-HCl, 150 Mm of NaCl, 0.05% Tween 20 and pH 7.2). The NC membrane was blotted with 5% non-fat milk in TBST for 1 h, followed by incubation with a primary Ab diluted with TBST for 2 h at 37°C. The membrane was washed with TBST three times and incubated with a secondary Ab at 37°C for 2 h. After washing with PBS three times, the NC membrane was stained using a HRP-DAB kit (Zhongshan Golden Bridge, China).
Abs used herein are as follows: primary Abs were rabbit anti-SUMO-2 mAb (Abcam, UK), rabbit anti-β-actin mAb (Jackson ImmunoResearch, USA); the secondary Ab was HRP-conjugated mouse anti-rabbit IgG (Bioworld, USA).
Abs used herein are as follows: primary Abs were rabbit anti-SUMO-2 mAb (Abcam, UK), rabbit anti-β-actin mAb (Jackson ImmunoResearch, USA); the secondary Ab was HRP-conjugated mouse anti-rabbit IgG (Bioworld, USA).
4
Comprehensive Cell and Tissue Analysis
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Quantitative real‐time polymerase chain reaction (qRT‐PCR) reagents and supplies are AG RNAex Pro RNA kit, SYBR Green Pro Taq HS kit, Evo M‐MLV reverse‐transcription kit (removal gDNA reagent) and Rox and were purchased from Accurate Biotechnology (Hunan) Co. Ltd. People's Republic of China. Western‐blotting reagents and supplies are Rabbit Anti‐Ngb, Polyclonal Antibody (bs‐1859R), Rabbit Anti‐HIF‐1, Alpha Polyclonal Antibody (bs‐0737R), Rabbit Anti‐beta‐Actin (Loading Control), Polyclonal antibody (bs‐0737R) and goat anti‐rabbit IgG/HRP(bs‐0295G‐HRP) and were purchased from Bioss Co. Ltd. People's Republic of China. RIPA tissue or cell rapid lysate was purchased from Bio topped, and 0.22μm polyvinylidene difluoride filter (PVDF) membranes, 4 × protein loading buffer (DTT), Rainbow 245 broad‐spectrum protein marker (11‐245KD) and ECL hypersensitivity luminescent solution were purchased from Solarbio Co. Ltd. People's Republic of China. Immunohistochemical reagents and supplies are immunohistochemical staining kit and HRP‐DAB kit and were purchased from Beijing Zhongshan Golden Bridge Biotechnology Co. Ltd. People's Republic of China.
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