For perform immunohistochemical (IHC) staining for RNF2, we purchased a tissue microarray slide from Servicebio (LVC-1608; Servicebio, consisting of 83 pairs of hepatocellular carcinoma tissue and adjacent normal tissues). Tumor tissue sections from nude mice were processed according to standard protocols and routinely examined by H&E staining. Firstly, the tissue sections were heated at 60 °C for 2 h, and then dewaxed with an alcohol gradient treatment. Then blocking the slides with 3% normal sheep serum (ZSbio, Beijing, China) for 1 h at room temperature. After blocking, specimens will be incubated overnight at 4 °C with primary antibody. The VECTASTAIN Elite ABC HRP kit (Vector Laboratories, Burlingame, CA, USA) and the VECTOR DAB kit (Vector Laboratories) were then used to color development according to the manufacturer’s instructions. The slides were visualized using Pannoramic Digital Slide Scanners (3DHISTECH, Budapest, Hungary).
Normal sheep serum
Manufactured by ZSGB-BIO
Sourced in China
Normal sheep serum is a biological fluid obtained from the blood of healthy sheep. It contains a complex mixture of proteins, electrolytes, and other biomolecules that are naturally present in the sheep's circulatory system.
Automatically generated - may contain errors
Lab products found in correlation
Vector dab kit, by Vector Laboratories (2 mentions)
Pannoramic digital slide scanner, by 3DHISTECH (2 mentions)
Vectastain elite abc hrp kit, by Vector Laboratories (2 mentions)
Fluoroshield mounting medium with dapi, by Abcam (2 mentions)
Multi channel fluorescence microscope, by Nikon (2 mentions)
Gapdh mouse monoclonal antibody, by Proteintech (1 mentions)
Anti gsdmd antibody, by Abcam (1 mentions)
Coralite594 conjugated donkey anti rabbit igg h l, by Proteintech (1 mentions)
Tunel apoptosis assay kit, by Keygen Biotech (1 mentions)
Triton x 100, by Keygen Biotech (1 mentions)
Cyclophosphamide, by Baxter (1 mentions)
5 protocols using normal sheep serum
1
Immunohistochemical Staining of RNF2 in Hepatocellular Carcinoma
2
Ovarian Dysfunction Mechanism Exploration
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The chemicals and reagents used were the standard substances kaempferol, quercetin, hesperetin, and hesperidin (National Institutes for Food and Drug Control), Cyclophosphamide (Baxter Oncology GmbH, Halle, Germany; No. 8K274A), Dehydroepiandrosterone (General Nutrition Corporation Pittsburgh, PA, USA; No. 61411H18), Rat FSH enzyme-linked immunosorbent assay (ELISA) kit (Cloud-Clone Corp. Wuhan, China; No. CEA830Ra), Rat AMH ELISA kit (Cloud-Clone Corp. Wuhan, China; No. CEA228Ra), Rat GnRH ELISA kit (Cloud-Clone Corp. Wuhan, China; No. CEA843Ra), Rat E2 ELISA kit (Cloud-Clone Corp. Wuhan, China; No. CEA461Ge), TUNEL apoptosis assay kit (KeyGENBio TECH. Jiangsu, China; No. KGA 7072), 4′, 6-diamidino-2-phenylindole (DAPI) staining kit (KeyGENBio TECH. Jiangsu, China; No. KGA 215-10), Normal sheep serum (ZSGB-BIO. Beijing, China; No. ZLI-9022), Triton X-100 (KeyGENBio TECH. Jiangsu, China; No. KGF011), Caspase-1 rabbit polyclonal antibody (Proteintech Group, Inc. Chicago, USA; No. 22915-1-AP), GAPDH mouse monoclonal antibody (Proteintech Group, Inc. Chicago, USA; No. 60004-1-lg), Anti-GSDMD antibody (Abcam Cambridge, MA, USA; No. ab219800), Rat IL-18 ELISA kit (RayBiotech, Inc. Georgia, USA; No. P97636), and CoraLite594-conjugated donkey anti-rabbit IgG (H+L) (Proteintech Group, Inc. Chicago, USA; No. SA00013-8).
3
Immunofluorescence Assay for Protein Localization
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Cells were inoculated with 24-well plates and cultured overnight, then fixed with formaldehyde and permeabilized with 0.1% Triton X-100. Blocking cells with 3% normal sheep serum (ZSbio, Beijing, China), then cells will be incubated with primary antibodies (DHX9, 1:200; TGFβR1, 1:200) overnight at 4°C. Next day, after incubated the fluorophore-conjugated secondary antibody, cell nuclei were be labelled by Fluoroshield mounting medium with DAPI (Abcam). Fluorescence images are acquired by a multi-channel fluorescence microscope (Nikon, Japan).
4
Immunohistochemical Analysis of Tumor Specimens
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All tissue specimens involved in this study were freshly obtained from patients undergoing surgical treatment at Xiangya Hospital, Central South University. Ethical approval was approved by the ethics committee of Xiangya Hospital, Central South University.
Tumor tissue sections were processed according to standard protocols. After heated and dewaxed with an alcohol gradient treatment, slides were blocked with 3% normal sheep serum (ZSbio, Beijing, China) for 1 h at room temperature. Then, specimens will be incubated overnight at 4°C with primary antibody (DHX9, 1:100; TGFβR1, 1:100). The VECTASTAIN Elite ABC HRP kit (Vector Laboratories, Burlingame, CA, United States) and the VECTOR DAB kit (Vector Laboratories) were then used to color development according to the manufacturer’s instructions. The slides were visualized using Pannoramic Digital Slide Scanners (3DHISTECH, Budapest, Hungary).
Tumor tissue sections were processed according to standard protocols. After heated and dewaxed with an alcohol gradient treatment, slides were blocked with 3% normal sheep serum (ZSbio, Beijing, China) for 1 h at room temperature. Then, specimens will be incubated overnight at 4°C with primary antibody (DHX9, 1:100; TGFβR1, 1:100). The VECTASTAIN Elite ABC HRP kit (Vector Laboratories, Burlingame, CA, United States) and the VECTOR DAB kit (Vector Laboratories) were then used to color development according to the manufacturer’s instructions. The slides were visualized using Pannoramic Digital Slide Scanners (3DHISTECH, Budapest, Hungary).
5
Immunofluorescence Staining of Cultured Cells
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Cells to be tested were inoculated with 24-well plates and cultured overnight, then fixed with formaldehyde at −20 °C for 20 min and permeabilized with 0.1% Triton X-100. Blocking cells with 3% normal sheep serum (ZSbio, Beijing, China) for 1 h at room temperature, then cells will be incubated with primary antibodies overnight at 4 °C. Next day, incubate the fluorophore-conjugated secondary antibody at room temperature. Fluoroshield mounting medium with DAPI (Abcam) was used for nuclei labeling. Fluorescence images are acquired by a multi-channel fluorescence microscope (Nikon, Japan).
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