Total protein was extracted via RIPA lysis (Beyotime, Shanghai, China) with phenyl-methane-sulfonyl fluoride and protease inhibitor. 10% SDS-PAGE gel electrophoresis with 30 μg protein per well was performed and then we transferred PVDF membranes. Antibodies PRPF6 (No. ab99292, 1:2000, Abcam, London, England), CEBPB (No. ab32358, 1:2000, Abcam, London, England), GATA3 (No. ab199428, 1:2000, Abcam, London, England), Vimentin (No. ab40497, 1:1000, Elabscience, Wuhan, China), E-cadherin (No. ab40285, 1:1000, Elabscience, Wuhan, China), N-cadherin (No. ab70061, 1:1000, Elabscience, Wuhan, China), β-tubulin III (No. YM0634, 1:2000, Immunoway, Wuhan, China), β-actin (No. AP0060, 1:5000, Bioworld, Wuhan, China) were incubated overnight at 4°C.
Prpf6
Manufactured by Abcam
Sourced in United Kingdom
PRPF6 is a laboratory product that is part of the U4/U6 small nuclear ribonucleoprotein (snRNP) complex, which plays a role in pre-mRNA splicing. The product is used for research purposes to study the function and structure of this protein.
Automatically generated - may contain errors
Lab products found in correlation
Cebpb, by Abcam (9 mentions)
Gata3, by Abcam (6 mentions)
Ripa lysis, by Beyotime (3 mentions)
β actin, by Bioworld Technology (3 mentions)
N cadherin, by Elabscience (3 mentions)
Vimentin, by Elabscience (3 mentions)
β tubulin 3, by Immunoway (3 mentions)
E cadherin, by Elabscience (3 mentions)
Magna rip kit, by Merck Group (3 mentions)
Fluorescent labelled secondary antibody, by Proteintech (3 mentions)
9 protocols using prpf6
1
Protein Extraction and Western Blot Analysis
2
Magna RIP-Seq Profiling of SNHG16
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The experiment was conducted followed the manufacturer’s protocol of Magna RIP Kit (Millipore, Massachusetts, USA). The antibodies were PRPF6 (10 ug per reaction, Abcam, London, England) and CEBPB (10 ug per reaction, Abcam, London, England). RNA was extracted after detachment from the bead using protease K. The expression of SNHG16 pre-mRNA, SNHG16-L and SNHG16-S was determined using RT-qPCR.
3
Immunofluorescence Staining of Proteins
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Cells growing on coverslips in 6-well plates were removed and fixed with 4% paraformaldehyde. Then, permeabilized in 0.5%–1.0% Triton X-100 for 10 min and blocked with 5% BSA for 30 min. The cells were then incubated with antibodies PRPF6 (1:150, Abcam, London, England), CEBPB (1:150, Abcam, London, England), GATA3 (1:150, Abcam, London, England) overnight at 4°C. Fluorescent-labelled secondary antibody (1:100, Proteintech, Wuhan, China) was added and incubated in the dark for 2 h. Cells were stained with DAPI and observed under the fluorescent microscope.
4
Protein Expression Analysis by Western Blot
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Total protein was extracted via RIPA lysis (Beyotime) with phenyl-methane-sulfonyl uoride and protease inhibitor. 10% SDS-PAGE gel electrophoresis with 30µg protein per well was performed and then we transferred PVDF membranes. Antibodies PRPF6 (1:2000, Abcam), CEBPB (1:2000, Abcam), GATA3(1:2000, Abcam), Vimentin (1:1000, Elabscience), E-cadherin (1:1000, Elabscience), N-cadherin (1:1000, Elabscience), β-tubulin III (1:2000, Immunoway), β-actin (1:5000, Bioworld) were incubated overnight at 4℃.
5
RIP Analysis of SNHG16 Isoforms
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The experiment was conducted followed the manufacturer's protocol of Magna RIP kit (Millipore). The antibodies used for RIP were PRPF6(10ug per reaction, Abcam) and CEBPB (10ug per reaction, Abcam).
RNA was extracted after detachment from the bead using protease K. The expression of SNHG16 pre-mRNA, SNHG16-L and SNHG16-S was determined using RT-qPCR.
RNA was extracted after detachment from the bead using protease K. The expression of SNHG16 pre-mRNA, SNHG16-L and SNHG16-S was determined using RT-qPCR.
6
Immunofluorescence Staining of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells growing on coverslips in 6-well plates were removed and xed with 4% paraformaldehyde. Then, permeabilized in 0.5-1.0% Triton X-100 for 10 min and blocked with 5% BSA for 30 minutes. The cells were then incubated with antibodies PRPF6 (1:150, Abcam), CEBPB (1:150, Abcam), GATA3(1:150, Abcam) overnight at 4℃. Fluorescent-labelled secondary antibody (1:100, Proteintech) was added and incubated in the dark for 2h. Cells were stained with DAPI and observed under the uorescent microscope.
7
Western Blot Analysis of Protein Markers
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Total protein was extracted via RIPA lysis (Beyotime) with phenyl-methane-sulfonyl uoride and protease inhibitor. 10% SDS-PAGE gel electrophoresis with 30μg protein per well was performed and then we transferred PVDF membranes. Antibodies PRPF6 (1:2000, Abcam), CEBPB (1:2000, Abcam), GATA3(1:2000, Abcam), Vimentin (1:1000, Elabscience), E-cadherin (1:1000, Elabscience), N-cadherin (1:1000, Elabscience), β-tubulin III (1:2000, Immunoway), β-actin (1:5000, Bioworld) were incubated overnight at 4℃.
8
RNA Extraction and Expression Analysis
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The experiment was conducted followed the manufacturer's protocol of Magna RIP kit (Millipore). The antibodies were PRPF6(10ug per reaction, Abcam) and CEBPB (10ug per reaction, Abcam). RNA was extracted after detachment from the bead using protease K. The expression of SNHG16 pre-mRNA, SNHG16-L and SNHG16-S was determined using RT-qPCR.
9
Immunofluorescence Staining of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells growing on coverslips in 6-well plates were removed and xed with 4% paraformaldehyde. Then, permeabilized in 0.5-1.0% Triton X-100 for 10 min and blocked with 5% BSA for 30 minutes. The cells were then incubated with antibodies PRPF6 (1:150, Abcam), CEBPB (1:150, Abcam), GATA3(1:150, Abcam) overnight at 4℃. Fluorescent-labelled secondary antibody (1:100, Proteintech) was added and incubated in the dark for 2h. Cells were stained with DAPI and observed under the uorescent microscope.
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