Tissues were fixed with 4% paraformaldehyde solution for at least 4 h at room temperature, followed by dehydration, dipping in wax, paraffin embedding and cut into sections. Then, these sections were treated with HE staining. For Immunohistochemistry, sections were incubated with serum or BSA for 30 min at room temperature, and then were dipped in diluted primary antibody for 2 h and then incubated with secondary antibody. The antibodies used in this research: primary antibodies: Proteintech (Wuhan, China): anti-SSRP1 (1:200; 15696-1-AP), anti-BCL2 (1:200; 26593-1-AP), anti-BAX (1:200; 50599-2-Ig), anti-MMP2 (1:200; 10373-2-AP), anti-MMP9 (1:200; 10375-2-AP) and PCNA (1:50; SC-56) from Santa Cruz Biotechnology; secondary antibody: goat anti-Rabbit IgG (H+L) (1:200; SA0000I-2) from Proteintech Group Inc. The sample was observed under BX53 fluorescence microscope (Olympus, Japan). Cells stained brown were positive cells.
Anti ssrp1
Manufactured by Proteintech
Sourced in China
Anti-SSRP1 is a protein that binds to the SSRP1 subunit of the FACT (facilitates chromatin transcription) complex. The FACT complex plays a role in regulating chromatin structure and gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit igg h l, by Proteintech (2 mentions)
Bx53 fluorescence microscope, by Olympus (1 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Sc 56, by Santa Cruz Biotechnology (1 mentions)
Anti mmp 9, by Santa Cruz Biotechnology (1 mentions)
Anti mmp2, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Anti p akt, by Proteintech (1 mentions)
Anti bcl2, by Arigo Biolaboratories (1 mentions)
Anti bax, by Arigo Biolaboratories (1 mentions)
Anti β actin, by Arigo Biolaboratories (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti gsk3β, by Proteintech (1 mentions)
Anti p gsk3β ser9, by Proteintech (1 mentions)
Anti lamin b1, by Proteintech (1 mentions)
Anti p21, by Proteintech (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Anti β catenin, by Proteintech (1 mentions)
Anti cyclin d1, by Proteintech (1 mentions)
3 protocols using anti ssrp1
1
Immunohistochemical Analysis of Tissue Samples
2
Western Blot Analysis of Protein Expression
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Cells and tissues were harvested and lysed in RIPA buffer for 30 min at 4-8ºC. Proteins were detected and quantified using BCA protein assay kit (Thermo Fisher Scientific). Protein samples (30-50 μg) were separated by 12% SDS-PAGE and transferred onto polyvinylidene fluoride membranes. Nonspecific binding sites were blocked with 5% low-fat milk and 0.1% Tween-20 at room temperature for 2 h. Subsequently, the membranes were incubated overnight at 4°C with the following primary antibodies: Proteintech (Wuhan, China): anti-SSRP1 (1:1000; 15696-1-AP), anti-p27 (1:1000; 25614-1-AP), anti-p21 (1:1000; 10355-1-AP), anti-CyclinD1 (1:1000; 60186-1-Ig), anti-BCL2 (1:1000; 26593-1-AP), anti-BAX (1:1000; 50599-2-Ig), anti-MDM2 (1:1000; 19058-1-AP), anti-p53 (1:1000; 10442-1-AP), anti-AKT (1:1000; 60203-2-Ig), anti-β-actin (1:2000; 66009-1-Ig) and arigo (Taiwan, China): anti-P-AKT (1:1000; ARG51559); secondary antibody: goat anti-Mouse (1:2000; 10828-I-AP) and goat anti-Rabbit IgG (H+L) (1:2000; SA0000I-2) from Proteintech Group Inc. The membranes were scanned for statistical analysis by enhanced chemiluminescence (ECL) using a gel image processing system (Tanon, Shanghai) and the density of the bands was analyzed using Image J ( Wayne Rasband National Institutes of Health, USA).
3
Western Blot Analysis of Cell Signaling Proteins
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Cells and tissues were harvested and lysed in RIPA buffer for 30 min at 4-8 °C. Proteins were detected and quanti ed with a BCA protein assay kit (Thermo Fisher Scienti c, Inc., MA, USA). Protein samples (30-50 μg) were separated with 12% SDS-PAGE and transferred to polyvinylidene uoride membranes.
Nonspeci c binding was blocked with 5% low-fat milk with Tween-20 for 2 h at room temperature. Subsequently, the membranes were incubated overnight at 4 °C with the following primary antibodies:
anti-SSRP1 (Proteintech, Chicago, IL, USA), anti-p21 (Proteintech, Chicago, IL, USA), anti-CyclinD1 (Proteintech, Chicago, IL, USA), anti-GSK3β (Proteintech, Chicago, IL, USA), anti-p-GSK3β (Ser9) (Proteintech, Chicago, IL, USA), anti-β-catenin (Proteintech, Chicago, IL, USA), anti-Lamin B1 (Proteintech, Chicago, IL, USA), anti-β-actin (Proteintech, Chicago, IL, USA) and anti-cleaved caspase3 (Cell Signaling Technology, Beverly, Massachusetts, USA). Lamin B1, encoded by the LMNB1 gene, is composed of a two-dimensional matrix of proteins located next to the inner nuclear membrane, which has been highly conserved during evolution. Therefore, Lamin B1 is often used as a nuclear protein internal control. The membranes were scanned for statistical analysis by enhanced chemiluminescence with a gel image processing system (Tanon, Shanghai, China).
Nonspeci c binding was blocked with 5% low-fat milk with Tween-20 for 2 h at room temperature. Subsequently, the membranes were incubated overnight at 4 °C with the following primary antibodies:
anti-SSRP1 (Proteintech, Chicago, IL, USA), anti-p21 (Proteintech, Chicago, IL, USA), anti-CyclinD1 (Proteintech, Chicago, IL, USA), anti-GSK3β (Proteintech, Chicago, IL, USA), anti-p-GSK3β (Ser9) (Proteintech, Chicago, IL, USA), anti-β-catenin (Proteintech, Chicago, IL, USA), anti-Lamin B1 (Proteintech, Chicago, IL, USA), anti-β-actin (Proteintech, Chicago, IL, USA) and anti-cleaved caspase3 (Cell Signaling Technology, Beverly, Massachusetts, USA). Lamin B1, encoded by the LMNB1 gene, is composed of a two-dimensional matrix of proteins located next to the inner nuclear membrane, which has been highly conserved during evolution. Therefore, Lamin B1 is often used as a nuclear protein internal control. The membranes were scanned for statistical analysis by enhanced chemiluminescence with a gel image processing system (Tanon, Shanghai, China).
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