Tumor-bearing brain tissue were collected from mice of each group. Tissues were fixed in 10% buffered formalin for 24 h following standard procedure for processing, paraffin-embedding, and sectioning to assess PCNA (1:200, Servicebio, GB11010), E-cadherin (1:200, Servicebio, GB14076) by IHC assay. The reaction was visualized using the Servicebio image analysis system, the staining was scored by two independent and experienced pathologists and calculated as the product of the staining intensity. First divides the positive grade: negative without staining, score 0, weak positive light yellow, score 1, medium positive brown, score 2, strong positive brown, score 3 points. Then analyze and calculate the area of weak, medium, and strong positive in the measurement area, the tissue area of the measurement area, the cumulative optical density value of the positive and the positive area. PCNA and E-cadherin quantification for each sample was determined and molecular data using modified H-scores ([{% of weak staining} × 1] + [{% of moderate staining} × 2] + [{% of strong staining} × 3]), to determine the overall percentage of PCNA and E-cad positivity across the entire stained sample, yielding a range from 0 to 300 [22 (link), 23 (link)].
Gb11010
Manufactured by Wuhan Servicebio Technology
Sourced in China
The GB11010 is a piece of laboratory equipment. It is designed to perform a specific function within a laboratory setting. However, a detailed and unbiased description of its core function cannot be provided without the risk of extrapolation or interpretation. Therefore, the description for this product is not available.
Automatically generated - may contain errors
Lab products found in correlation
Gb11001, by Wuhan Servicebio Technology (2 mentions)
E cadherin, by Wuhan Servicebio Technology (1 mentions)
Microscopy and imaging system, by Olympus (1 mentions)
Af5330, by Affinity Biosciences (1 mentions)
Af6241, by Affinity Biosciences (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Af3241, by Affinity Biosciences (1 mentions)
Gb14078, by Wuhan Servicebio Technology (1 mentions)
Cd79a, by Novus Biologicals (1 mentions)
Af7001, by Abcam (1 mentions)
Epcam, by Wuhan Servicebio Technology (1 mentions)
Anti pcna antibody, by Wuhan Servicebio Technology (1 mentions)
P0015, by Beyotime (1 mentions)
Nonfat milk, by BD (1 mentions)
Gb11226, by Wuhan Servicebio Technology (1 mentions)
Gb23303, by Wuhan Servicebio Technology (1 mentions)
Total protein extraction kit, by BestBio (1 mentions)
Gb111499, by Wuhan Servicebio Technology (1 mentions)
Wla019, by Wanlei (1 mentions)
A3107, by ABclonal (1 mentions)
8 protocols using gb11010
1
Immunohistochemical Analysis of PCNA and E-cadherin
2
Immunohistochemical Localization of Gene Expression in Uterovaginal Junction Tissue
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Immunohistochemistry was conducted to localize the expression of genes of interest in UVJ tissue samples. The UVJ tissues fixed in PFA were dehydrated, embedded with wax, and processed sections with 5 μm thickness. The sections were gone through dewaxing with xylene, antigen retrieval with sodium citrate buffer (pH 6.0 ± 0.1), subsequently incubated with specific primary antibodies (GNAQ, rabbit, Thermo Fisher, PA5-79318, 1:2000; ST6GAL1, mouse, Abcam, ab77676, 1:200, UK; proliferating cell nuclear antigen [PCNA ], rabbit, Servicebio, GB11010, 1:2000, China; adipocyte differentiation-related protein [ADFP ], rabbit, Huabio, ET1704-17, 1:1000, China) overnight at 4°C. The sections were then incubated with secondary antibody (Proteintech, China) for 1 h at room temperature and stained by using DAB (1:50) followed with hematoxylin counterstain. The stained sections were sealed and recorded as images with microscopy and imaging system (Olympus, Japan). The experiments were performed at least twice in 2 individuals each group per antibody staining.
3
Protein Expression Analysis in LAC Cells
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LAC tissue and cell lines were lysed with RIPA buffer. The supernatants were resolved in SDS-PAGE and transferred onto polyvinylidene fluoride membranes (IPVH00010, Millipore, MA, USA), which were then probed with anti-HNRNPA2B1 (Proteintech, 1:200, Lot No. 67445-1-Ig, Wuhan, China), anti-PTEN (Proteintech, 1:1000, 60300-1-Ig, Wuhan, China), anti-PI3K (1:1000, AF6241, Affinity), anti-p-PI3K (1:1000, AF3241, Affinity), anti-AKT (Proteintech, 1:1000, 10176-2-AP, Wuhan, China), anti-p-AKT (Proteintech, 1:1000, 66444-1-Ig, Wuhan, China), anti-PCNA (1:1000, GB11010, Servicebio), anti-MMP2 (1:1000, AF5330, Affinity) and anti-GAPDH (1:1000, AB-P-R 001, GOODHERE, Hangzhou, China) overnight at 4 °C. Protein bands were emerged by enhanced chemiluminescence method.
4
Tissue Microarray Analysis of Lung Adenocarcinoma
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We established a tissue microarray (TMA, 3 mm) consisting of LUAD samples: 14 AIS samples, 18 MIA samples, and 17 IAC samples (Supplementary Table 4 ). After antigen repair and blocking, the TMA was incubated with specific primary antibodies (EPCAM, Servicebio, GB14078, 1:200; FOXP3, Servicebio, GB11093, 1:200; TPSB2, Novus, NBP2-33551, 1:300; CD79A, Novus, NB100-64347ss, 1:200; CLDN5, Servicebio, GB11290, 1:200; COL1A1, Affinity, AF7001, 1:200; UBE2C, Abcam, ab12290, 1:50; p-SMAD2, Affinity, AF8314, 1:200; SCGB1A1, Servicebio, GB111412, 1:200; PDPN, Affinity, AF3670-SP, 1:200; FCN1, Novus, NBP1-84706, 1:200; GHD, Proteintech, 67538 1:250; PCNA, Servicebio, GB11010, 1:300), incubated with horseradish peroxidase (HRP)-labeled secondary antibodies or fluorophore-labeled secondary antibodies, and finally stained with diaminobenzidine and counterstained with hematoxylin or stained with DAPI. Three independent pathologists distinguished the pathological type for the TMA.
5
Schwann Cell Protein Expression Analysis
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The Schwann cells were co-cultured with BDNF for 3 days and then the cells were harvested for western blot analysis. And then the Schwann cells protein extracts were prepared by Total Protein Extraction Kit (BestBio, China). All the cell proteins were mixed with sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) loading buffer (P0015, Beyotime) and heated at 100°C for 5 min. Protein samples (80 μg/well) were loaded on SDS–PAGE (10–12%) and electroplated onto polyvinylidene fluoride (PVDF) films (Millipore, United States). The PVDF films were blocked with 5% non-fat milk (BD, United States) for 60 min at room temperature and subsequently incubated overnight at 4°C with the primary antibodies: anti-PCNA antibody (GB11010, Servicebio), anti-MBP antibody (GB11226, Servicebio), and rabbit β-actin Rabbit antibody (GB11001, Servicebio). After incubating the PVDF films with horseradish peroxidase (HRP)-labeled secondary antibodies (GB23303, Servicebio) for 60 min at 25°C, the signal was collected by Image Studio Digits Ver 4.0. Density values were normalized to β-actin. The quantification of Western blot data was performed using Image-J software.
6
Cancer Cell Proliferation Analysis
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7
Chondrocyte Protein Extraction and Western Blot
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The total protein or nuclear protein of chondrocytes was extracted using a protein extraction kit (WLA019, Wanleibio, Shenyang, China) and a nuclear protein extraction kit (P0028, Beyotime, Shanghai, China). The protein concentration was assessed by BCA kit (G2026, Servicebio, Wuhan, China). Forty-gram proteins per sample were separated by 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes. The membranes were blocked for 2 h with 5% skim milk, and incubation with anti-BAX (1:2000, 50599, Proteintech, Wuhan, China), anti-BCL2 (1:500, A0208, Abclonal, Wuhan, China), anti-β-ACTIN (1:2000, GB11001, Servicebio, Wuhan, China), anti-NFAT1 (1:1000, A3107, Abclonal), and anti-PCNA (1:1000, GB11010, Servicebio, Wuhan, China) at 4 °C was performed overnight. After incubation with HRP-conjugated secondary antibodies, target bands were visualized using the ChemiDoc Touch (Bio-Rad, CA, USA).
8
Histopathological Analysis of Stomach Specimens
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Stomach specimens were dissected along the greater curvature. After washing with phosphate buffer saline, specimens were fixed in paraformaldehyde, embedded in paraffin, and sliced into 4 μm sections. Each sample was stained with haematoxylin–eosin (H&E) (G1005, Servicebio, Wuhan, China) and Masson trichrome (G1006, Servicebio, Wuhan, China) according to the manufacturer s instructions. Moreover, terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) (In Situ Cell Death Detection Kit, Roche, Basel, Switzerland) and the expression of E-cadherin (1:200) (GB12082, Servicebio, Wuhan, China), β-Catenin (1:200) (G12015, Servicebio, Wuhan, China), CD117 (1:2000) (GB11073-2, Servicebio, Wuhan, China), PGP9.5 (1:100) (GB11159-1, Servicebio, Wuhan, China), and proliferating cell nuclear antigen (PCNA) (1:100) (GB11010, Servicebio, Wuhan, China) were detected by immunofluorescence following the manufacturer’s instructions. Histopathological analysis was performed by two experienced pathologists unaware of the group allocation.
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