Briefly, the tissues that were previously formalin-fixed and paraffin-embedded were sliced into 4 µm sections, and were then incubated with the primary antibody recognizing human DAB2IP (#ab87811, abcam) at 1:200 dilution at 4°C overnight. The protein was visualized using a tissue staining kit (Zhongshan Biotechnology, Beijing, China) according to the manufacturer’s instructions. The final staining score was determined by color intensity and positive cell rate, ranging 0–12 which was described in our previous study.23 (link),24 (link) Patients were classified into two groups: scores 0–4 were considered as none or low, while 5–12 were considered as high expression.
Ab87811
Manufactured by Abcam
Sourced in United Kingdom
Ab87811 is a laboratory instrument for the separation and purification of biological molecules. It utilizes a technique called high-performance liquid chromatography (HPLC) to efficiently isolate and concentrate target analytes from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Tissue staining kit, by Zhongshan Biotechnology (1 mentions)
Cocktails of protease and phosphatase inhibitors, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Mouse monoclonal anti β actin ab, by Merck Group (1 mentions)
Peroxidase conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Vazyme (1 mentions)
4 protocols using ab87811
1
Immunohistochemical Analysis of DAB2IP
2
Western Blot Analysis of Signaling Pathways
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Cells were lysed in RIPA lysis buffer (Beyotime Institute of Biotechnology) supplemented with cocktails of protease and phosphatase inhibitors (Sigma). A total of 20 µg protein was then separated by 8% SDS‑PAGE and then transferred onto a PVDF membranes according to methods described previously.23 (link),24 (link) Antibodies against DAB2IP (dilution 1:1000; #ab87811, abcam), cyclin D1 (dilution 1:1000; 26939-1-AP, proteintech), p21 (dilution 1:1000; 10355-1-AP, proteintech), p-AKT (Ser473) (dilution 1:1000; #4058, CST), AKT (dilution 1:1000; #9272, CST), p-ERK (dilution 1:1000; #4370, CST), ERK (dilution 1:1000; #4695, CST) and Tubulin (dilution 1:2000; #AF5012, Beyotime) were used in this study.
3
Immunoblotting for Chromatin Proteins
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The following antibodies were used: a mouse monoclonal Ab anti-EZH2 (clone 144CT2.1.1.5) (Thermo Fisher Scientific, Rockford, USA) (1:250), a rabbit Polyclonal anti-DAB2IP Ab (ab87811, Abcam, Cambridge, UK) (1:500), a rabbit polyclonal anti-H3K27me3Ab (Millipore, CA, USA)(1:250), a mouse monoclonal anti-β-Actin Ab (Sigma, St. Louis, MO) (1:20,000), and a mouse Monoclonal anti-Vimentin Ab (Clone Vim 3B4, Dakocytomation) (1:100).
4
Apoptosis and DNA Damage Signaling Assays
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Primary antibodies used in this study: rabbit anti-DAB2IP (ab87811, Abcam, UK); rabbit anti-cleaved poly (ADP-ribose) polymerase (PARP, Asp214), rabbit anti-cleaved caspase-3 (Asp175), mouse anti-Phospho-Histone H2AX (γ-H2AX) (Ser139), rabbit anti-53BP1, rabbit anti-AKT and anti-Phospho-AKT (Ser473), rabbit anti-JNK and anti-Phospho-JNK, rabbit anti-ERK1/2 and anti-Phospho-ERK1/2, rabbit anti-ASK1 and anti-Phospho-ASK1 (ser966), rabbit anti-14-3-3, rabbit anti-Thioredoxin and α-tubulin (Cell Signaling Technology, Boston. MA). Secondary antibodies: Dylight 549-conjugated goat anti-rabbit IgG (Proteintech Group, Inc., Chicago, IL), Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, Carlsbad, CA). Peroxidase-conjugated goat anti-rabbit IgG and goat anti-mouse IgG (Santa Cruz Biotechnology, Santa Cruz, CA). Annexin V-FITC/PI apoptosis detection kit (Vazyme Biotech, China).
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