Bacterial concentrations of cultured N. gonorrhoeae FA1090 and FA19 as well as two clinical strains were adjusted to 0.5 McFarland units with GCBL. Take 1 mL of the above bacterial solution and transferred to a centrifuge tube at 4 °C and 12000 rpm for 2 min, the supernatant was discarded, 1 × loading buffer was added, and the samples were boiled at 100 °C for 15 min for later use. Whole bacterial lysates were separated by SDS-PAGE and transferred to PVDF membranes. Western blotting was performed using anti-passenger or anti-translocator antisera as primary antibodies (1:1000), and HRP-labeled goat anti-mouse IgG (ZSGB-Bio, China) was used as a secondary antibody (1:5000) to analyze the expression of App in different N. gonorrhoeae.
Hrp labeled goat anti mouse igg
Manufactured by ZSGB-BIO
Sourced in China
HRP-labeled goat anti-mouse IgG is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to bind to mouse immunoglobulin G (IgG) and can be used in various immunoassay techniques, such as ELISA, Western blotting, and immunohistochemistry, to detect and quantify the presence of mouse IgG in samples.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Mouse anti mast cell tryptase antibody, by Abcam (1 mentions)
Diaminobenzidine dab kit, by ZSGB-BIO (1 mentions)
Aminopterin, by Merck Group (1 mentions)
Hypoxanthine, by Merck Group (1 mentions)
Rpmi 1640 medium powder, by Thermo Fisher Scientific (1 mentions)
Anti flag tag antibody, by Abnova (1 mentions)
β actin, by ZSGB-BIO (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Anti p stat3 tyr 705 antibody, by Cell Signaling Technology (1 mentions)
Goat anti rabbit igg, by ZSGB-BIO (1 mentions)
Polyethylene glycol 1450 solution peg 1450, by Merck Group (1 mentions)
7 protocols using hrp labeled goat anti mouse igg
1
Protein Expression Analysis of N. gonorrhoeae
2
Eosinophil and Mast Cell Degranulation Evaluation
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Immunohistochemistry (IHC) stains were used to evaluate the expression of major basic protein (MBP) and tryptase, which are markers of eosinophil and mast cell degranulation, respectively26 (link). After dewaxing and rehydrating, tissue sections (D1, D2) were treated with 0.3% hydrogen peroxide in methanol to block endogenous peroxidase activity followed by pepsin or heat-mediated antigen retrieval. After blocking with 3% goat serum for 20 minutes at room temperature to minimize nonspecific staining, the sections were incubated with mouse anti-eosinophil MBP antibody (1:50, AbD Serotec, UK) and mouse anti-mast cell tryptase antibody (1:26000, Abcam, USA) for 1 hour at room temperature. After washing, the sections were treated with HRP-labeled goat anti-mouse IgG (ZSGB, China) for 30 minutes. The reaction was visualized using diaminobenzidine (DAB kit, ZSGB, China) and hematoxylin staining. Eosinophil degranulation positive was defined by the presence of MBP granules stain in randomly selected fields. If there was evidence of eosinophil degranulation on IHC stain, five non-overlapping fields on the slides (400× magnification, Nikon) were randomly selected and analyzed by two independent observers (LD, LM), to determine the mean numbers of degranulated eosinophil and mast cell and expressed per HPF.
3
Monoclonal Antibody Production Protocol
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The anti-His6 antibody and HRP-labeled goat anti-mouse IgG were purchased from ZSGB-Bio Co., Ltd (Beijing, China). The control antibody LN22 (Newcastle, United Kingdom) and MaxVision/HRP IHC kit were provided by Fuzhou Maixin Biotech Co., Ltd (Fuzhou, China). SP2/0 myeloma cells were stored in our laboratory. Hypoxanthine, aminopterin and thymidine supplement (HAT), Hypoxanthine and thymidine supplement (HT) were purchased from Sigma-Aldrich (Shanghai, China). RPMI 1640 medium powder and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Shanghai, China). Balb/c mice were from Wushi Animal Laboratory (Shanghai, China). All animal studies were approved by the Animal Ethics Committee of the Fujian Agriculture and Forestry University and have been carried out in accordance with the NIH guidelines for the care and use of Laboratory animals.
4
Western Blot Analysis of p21Ras Protein
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WB assay was performed by extracting protein from cell samples with RIPA cellular lysate (Solarbio), followed by electrophoresis on SDS-PAGE gels and transfer of proteins to polyvinylidene fluoride (PVDF) membranes. Next, the PVDF membranes were incubated with BR2-p21Ras scFv (0.3mg/ml,dilution1:800) after using skimmed milk powder as a blocking agent and then were incubated with the anti-FLAG tag antibody(Abnova,MAB9744,China) at a 1:1000 dilution and horseradish peroxidase (HRP) labeled goat anti-mouse IgG (ZSGB-Bio, ZB-5305, China) at 1:1000. After washing with TBST (50mMTris, 150mMNaCl, and 0.5%Tween-20)for three times, The PVDF membranes were then stained with DAB. β-Actin (ZSGB-Bio,TA-09,China) protein was used as an internal control.
5
Immunoblot Analysis of S. japonicum Antigens
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The S. japonicum adult worm antigen (SjAWA) and rSj29 protein were separated with 12% SDS-PAGE (15 mg/well), and were transferred to a nitrocellulose membrane (Millipore, USA). For the immunoblot analyses, McAbs against Sj29 from hybridomas culture supernatant were used as the primary antibody. HRPlabeled goat-anti-mouse IgG (1:80,000, ZSGB-Bio, China) was used as the secondary antibody. The protein-antibody complexes were visualized using chemiluminescent analysis (Pierce, USA).
6
Glioma Tissue Microarray and IHC Analysis
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The tissue microarray (TMA) collected 55 glioma cases, in which 2 cases were of non‐tumor (cortical dysplasia), 2 cases of Grade I glioma, 12 cases of Grade II glioma, 12 Grade III, and 27 cases of GBM (Grade IV) (Figure S1 and Table S1). The procedure of immunohistochemistry (IHC) was performed as previously described.35 Primary antibodies: Anti‐IKBKE antibody (Cell Signaling, USA), Anti‐pSTAT3 (Tyr705) antibody (Cell Signaling, USA) and Anti‐CD274 antibody (Abcam, USA) were incubated with a dilution of 1:100. Secondary antibodies: HRP‐labeled goat anti‐mouse IgG and goat anti‐rabbit IgG were obtained from ZSGB‐Bio, China. The IHC images were observed and acquired though a light microscope (OLYMPUS, Japan).
7
Monoclonal Antibody Production Protocol
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The anti-6 × His tag monoclonal antibody and HRP-labeled goat anti-mouse IgG were purchased from ZSGB-Bio Co., Ltd. (Beijing, China). The cocktail of LK2H10 and PHE5 antibodies, and MaxVisionTM/HRP IHC kit were provided by Fuzhou Maixin Biotech Co., Ltd. (Fuzhou, China). SP12 antibody was from Thermo Fisher Scientific (Shanghai, China). Hypersensitivity ECL chemiluminescence detection kit was purchased from Wuhan Sanying Co., Ltd. (Wuhan, China). Myeloma cells SP2/0 were stored in our laboratory. Hypoxanthine, aminopterin and thymidine supplement (HAT), hypoxanthine and thymidine supplement (HT) and polyethylene glycol 1450 solution (PEG 1450) were purchased from Sigma-Aldrich (Shanghai, China). RPMI medium 1640 powder and fetal bovine serum (FBS) were purchased from Thermo Fisher (Shanghai, China). Balb/c mice were from Wushi Animal Laboratory (Shanghai, China). QuickAntibody-Mouse5W adjuvant (Quickadjuvant) was purchased from Beijing Biodragon Immunotechnologies Co., Ltd. (Beijing, China). All animal experiments obeyed the protocols approved by the Animal Ethics Committee of the Fujian Agriculture and Forestry University.
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