Formalin‐fixed and paraffin‐embedded tissue sections were incubated with Ki67 primary antibody (dilution 1:1000; Proteintech), PCNA antibody (dilution 1:300; Proteintech), and SF3A3 (dilution 1:200; Abcam) overnight at 4 °C. Then, the sections were incubated with HRP-conjugated anti-rabbit IgG secondary antibody (dilution 1:300; Yeasen) for 2 h at room temperature. The slides were evaluated by two independent observers.
Pcna antibody
Manufactured by Proteintech
Sourced in United States, China
The PCNA antibody is a primary antibody designed to detect the Proliferating Cell Nuclear Antigen (PCNA) protein, which is a key component of the DNA replication machinery. PCNA plays a critical role in cell cycle progression and DNA repair processes. This antibody can be used to study the expression and localization of PCNA in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
P21 antibody, by Abcam (2 mentions)
Ki67 antibody, by Abways (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Sf3a3, by Abcam (1 mentions)
Ki67 primary antibody, by Proteintech (1 mentions)
P53 antibody, by Proteintech (1 mentions)
Ki67 antibody, by Abcam (1 mentions)
Anti caspase 3 antibody, by Cell Signaling Technology (1 mentions)
E cadherin antibody, by Proteintech (1 mentions)
Vimentin antibody, by Proteintech (1 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Mmp 9 antibody, by Proteintech (1 mentions)
Bcl 2 antibody, by Proteintech (1 mentions)
Akt antibody, by Proteintech (1 mentions)
N cadherin antibody, by Proteintech (1 mentions)
P70 s6 kinase antibody, by Cell Signaling Technology (1 mentions)
Peroxidase conjugated secondary antibodies, by CWBIO (1 mentions)
Pakt ser473 antibody, by Cell Signaling Technology (1 mentions)
Rapamycin, by Meilun (1 mentions)
Bax antibody, by Proteintech (1 mentions)
7 protocols using pcna antibody
1
Immunohistochemical Analysis of Cell Proliferation
2
Evaluating Tumor Growth and Metastasis
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The protocol for animal experiments was viewed and approved by the Institutional Animal Care and Use Committee of Shanghai East Hospital, Tongji University. Four-week-old Male athymic nude mice were obtained from SLAC Laboratories Animal, Shanghai, China and maintained in sterile laminar flow cabinets. 1.5×106 of SGC7901 or MGC803 cells was inoculated bilaterally and subcutaneously into the flanks of nude mice. Bidimensional tumor measurements were taken with vernier calipers every 4 days, and the mice were euthanized after 4 weeks. The volume of the implanted tumor was calculated using the formula: volume = (π*width2*length)/6. In addition, the subcutaneous tissues were fixed in 4% paraformaldehyde, stored in 70% ethanol, and then treated with paraffin-embedding, sectioning, H&E staining and immunohistochemistry examination with PARI antibody (1:100, ABGENT, USA) and PCNA antibody (1:100, Proteintech, China).
For lung metastasis assay (n=6 for each group), the mice at the age of 6 weeks old were injected with 1×106 cells of SGC7901-Lv-shNC, SGC7901-Lv-shPARI through tail vein, respectively. After 4 weeks, the groups were sacrificed, the lungs were removed and photographed, obvious lung metastatic tumors on the surface were calculated.
For lung metastasis assay (n=6 for each group), the mice at the age of 6 weeks old were injected with 1×106 cells of SGC7901-Lv-shNC, SGC7901-Lv-shPARI through tail vein, respectively. After 4 weeks, the groups were sacrificed, the lungs were removed and photographed, obvious lung metastatic tumors on the surface were calculated.
3
Rapamycin and 5-FU Synergistic Effects on Colorectal Cancer
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Rapamycin and 5-FU were obtained from meilunbio (Dalian, China). The anti Caspase3 antibody, Cleaved Caspase3 antibody, p-AKT (ser 473) antibody, p-AKT (Thr 308) antibody, p70 S6 Kinase antibody, and p-p70S6 Kinase (Thr 389) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). The ki-67 antibody was purchased from Abcam (Cambridge, UK). The BCL-2 antibody, Bax antibody, AKT antibody, p53 antibody, E-Cadherin antibody, N-Cadherin antibody, Vimentin antibody, MMP-9 antibody and PCNA antibody were purchased from Proteintech (Wuhan, China). The GAPDH rabbit polyclonal antibody was obtained from XianZhi Biotech (Hangzhou, China). The peroxidase-conjugated secondary antibodies were obtained from CWBIO (Taizhou, China).
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
4
Immunofluorescence Analysis of Cell Senescence
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The tissue sections were deparaffinized and 3% H2O2 was used to block the endogenous peroxidase. The sections were incubated in tris-buffered saline (TBS) with 5% albumin bovine V (BSA; Solarbio, Beijing, China) for 1 h. The cell samples were washed with PBS three times and fixed with 4% paraformaldehyde for 15 min. After washing with PBS three times, tissue sections or cells were permeated with 0.3% Triton X-100 for 15 min and then incubated with p21 antibody (1:200; Abcam, Cambridge, MA, USA), p16 antibody (1:200; Cell Signaling Technology, Danvers, MA, USA), LC3 antibody (1:200; CST, Danvers, MA, USA), PCNA antibody (1:200; Proteintech, Wuhan, China) or Ki67 antibody (1:200; Abways, Shanghai, China) in 5% BSA overnight at 4 °C. After washing with TBS, the sections were incubated with a Rhodamine (TRITC)-conjugated goat anti-rabbit IgG (H+L) (1:300; Abcam, Cambridge, UK). The nuclei were counterstained with DAPI (Invitrogen, Waltham, MA, USA). The sections were washed with PBS three times, and coverslips were mounted in 90% glycerol in PBS. The fluorescence was detected by a fluorescence microscope (Nikon, Tokyo, Japan). Image J was used to analyze the fluorescence intensity.
5
Inhibition of Liver Cancer Growth
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Animal experiments were approved by the Animal Research Committee of Sun Yat-Sen University, and all procedures strictly followed the NIH Guide for Care and Use of Laboratory Animals. Female nude mice at age of 5-6 weeks were purchased from the Sun Yat-Sen University Laboratory Animal Center (Guangzhou, China). Approximately 5.0×106 SK-HEP-1/miR-1226-3p or SK-HEP-1/control cells were suspended in 100 μL PBS and injected subcutaneously into the right side of the posterior flank of female nude mice. When the average tumor size reached approximately 50 mm3, sorafenib (30 mg/kg) was administered via intraperitoneal injection every other day. All mice were killed after 2 weeks. The primary tumors were excised and analysed by immunohistochemistry of DUSP4 antibody (Proteintech Group, Wuhan, China) and PCNA antibody (Proteintech Group, Wuhan, China).
6
Adiponectin and TGF-β1 Modulate Vascular Remodeling
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Human adiponectin (Acrp30) and human TGF-β1 were purchased from Novoprotein (Beijing, China). Cell Counting Kit-8 (CCK-8) and EdU kits were procured from SolarBio (Beijing Solebo Technology Co., Ltd, Beijing, China). PCNA antibody, smooth muscle actin polyclonal (α-SMA) antibody, collagen type I (collagen I) rabbit polyclonal antibody, p38 MAPK antibody, p-p38 MAPK antibody, NF-κB antibody, p-NF-κB antibody, β-catenin antibody, p-β-catenin antibody, Notch1 antibody, and HRP-conjugated AffiniPure goat anti-mouse IgG (H + L) were obtained from Proteintech (Wuhan, China). SB203580 was purchased from MCE (Shanghai, China). Other chemicals used in this study, such as the BCA protein assay kit and SDS-PAGE gel separation kit, were acquired from ASPEN (ASPEN Biotechnology Co., Ltd, Wuhan, China).
7
Immunofluorescence Staining of Cellular Proteins
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Tissue sections were deparaffinized and endogenous peroxidase was blocked with 3% H2O2. Sections were incubated in 5% albumin bovine V (BSA, Solarbio) for 1 h. The cells were washed three times with PBS and fixed with 4% paraformaldehyde for 15 min. After three washes with PBS, the cells were permeated with 0.1% Triton X-100 for 15 min. After three washes with PBS, the cells were incubated with 5% bovine serum albumin for 30 min at room temperature and then incubated with p21 antibody (1:300, Abcam, Cambridge, MA, USA), p16 antibody (1:200, Abcam), SFTPC antibody (1:200, Abclonal, Wuhan, China), Keap1 antibody (1:300, Proteintech, Wuhan, China), Nrf2 antibody (1:300, Cell Signaling Technology, USA), PCNA antibody (1:200, Proteintech), Ki67 antibody (1:200, Abways, Shanghai, China), or Trim25 antibody (1:200, Proteintech) at 4 °C overnight. The next day, after washing with PBS, fluorescein-labeled secondary antibodies (1:300, Abcam) for immunofluorescence were incubated. Nuclei were counterstained with DAPI (Invitrogen, USA). The samples were washed three times with PBS and photographed with fluorescence microscopy (Nikon, Japan).
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