BCL1 and JVM-13 cells, after IBS and MBS treatment for 24 h, were fixed, permeabilized, and incubated with antibodies specific for Tyr 705 phosphorylated STAT3 (sc-8059, 200 µg/mL, Santa Cruz Biotech. Inc., Santa Crus CA, USA, dilution 1:300) and Ki-67 (11-5698-82, 100 µg/mL, eBioscience, San Diego, CA, USA, dilution 1:400). JVM-13 cell were stained with anti-cyclin D3 (ab28283, 100 µg/mL, Abcam Cambridge, UK, dilution 1:100) antibody. BCL1 cells were stained with anti-p21 (ab188224, 100 µL, Abcam Cambridge, UK, dilution 1:50), anti-p16 (ab211542, 100 µL, Abcam Cambridge, UK, dilution 1:500), and anti-27 antibodies (ab215434, 100 µL, Abcam Cambridge, UK, dilution 1:100). Cells were additionally incubated with secondary goat anti-mouse IgG FITC (ab6717-1, 1 mg/mL, Abcam Cambridge, UK, dilution 1:2000). The FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) was used for flow cytometric analysis, and the data were analyzed using FlowJo (Tree Star).
Ab215434
Manufactured by Abcam
Sourced in United Kingdom
Ab215434 is a laboratory equipment product from Abcam. It serves as a core functional tool for research applications. Details on its specific features and intended use are not provided to maintain an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (3 mentions)
Ab188224, by Abcam (3 mentions)
Ab211542, by Abcam (3 mentions)
Ab28283, by Abcam (3 mentions)
Permeabilization buffer, by BD (2 mentions)
Ic1799g, by Novus Biologicals (2 mentions)
Ma5 14336, by Thermo Fisher Scientific (2 mentions)
Ab150073, by Abcam (2 mentions)
Ab6785, by Abcam (2 mentions)
Sc 8059, by Santa Cruz Biotechnology (1 mentions)
Ab32087, by Abcam (1 mentions)
Ab13654, by Abcam (1 mentions)
Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions)
Ab68455, by Abcam (1 mentions)
Ab8227, by Abcam (1 mentions)
Ab2302, by Abcam (1 mentions)
4 protocols using ab215434
1
Evaluating Cell Signaling and Proliferation
2
Evaluating Cell Cycle Markers in 4T1 Cells
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4T1 cells, grown in culture plates, were treated with either DE-EDCP or cisplatin (31.25 μM) for 24 hours. Next, the treated cells were fixed and permeabilized with permeabilization buffer (BD Bioscience) and incubated with antibodies specific for STAT3 (IC1799G, Novus Biologicals, San Diego, USA), Ki-67 (11-5698-82, eBioscience, San Diego, USA), cyclin D3 (ab28283, Abcam Cambridge, United Kingdom), cyclin E (MA5-14336, Thermo fisher scientifics, USA), p16 (ab211542, Abcam Cambridge, United Kingdom), p21 (ab188224, Abcam Cambridge, United Kingdom) and p27 (ab215434, Abcam Cambridge, United Kingdom). For staining cyclin D3, cyclin E, p16, p21 and p27 cells were additionally incubated with secondary goat anti-mouse IgG FITC (ab6785 Abcam Cambridge, United Kingdom) or donkey anti-rabbit IgG (ab150073 Abcam Cambridge, United Kingdom). Flow cytometry was conducted on FACSCalibur flow cytometer (BD Biosciences, San Jose, USA) and the data were analyzed using FlowJo (Tree Star).
3
Investigating Apoptosis Pathway in BCL1 Cells
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BCL1 cells, grown in culture plates, were treated with or without Pt(S-pr-thiosal)2 (0.05 mg/mL) for 12 h. The treated cells were fixed and permeabilized with permeabilization buffer (BD Bioscience) and incubated with antibodies specific for Bcl-2 (11-6992-42, Thermo Fisher Scientific, Cambridge, UK), Noxa (ab13654, Abcam, Cambridge, UK), STAT3 (IC1799G, Novus Biologicals, San Diego, CA, USA), p16 (ab211542, Abcam, Cambridge, UK), p21 (ab188224, Abcam) and p27 (ab215434, Abcam), cyclin D3 (ab28283, Abcam Cambridge, UK), cyclin E (MA5-14336, Thermo fisher scientifics, Waltham, MA, USA), Ki-67 (151212, BioLegend, San Diego, CA, USA), Noxa (ab13654, Abcam) and Mcl-1 (ab32087, Abcam). For staining p16, p21, and p27 cells were additionally incubated with secondary goat anti-mouse IgG FITC (ab6785, Abcam) or donkey anti-rabbit IgG (ab150073, Abcam). Flow cytometry was performed on FACSCalibur flow cytometer (BD Biosciences). The data were analyzed using FlowJo (Tree Star).
4
Western Blot Analysis of Cell Signaling Proteins
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The cells were lysed and boiled at 96°C for 5 min, before loading onto four 20% SDS-polyacrylamide gels. Proteins were separated by electrophoresis in a Mini-PROTEAN Tetra cell chamber and transferred to polyvinylidene difluoride membranes. Then, the membranes were blocked in 5% non-fat milk (Yili Milk Company, China) in Tris-buffered saline-Tween (TBS-T) for 1 h at room temperature, and incubated with primary antibodies against Skp2 (ab68455, Abcam, UK), p27 (ab215434, Abcam), caspase-3 (ab2302, Abcam), and β-actin (ab8227, Abcam) overnight at 4°C with gentle shaking. Secondary antibodies conjugated to horseradish peroxidase (HRP) were applied for 1 h at room temperature, and immunoreactive bands were developed using enhanced chemiluminescence (Thermo Fisher Scientific, USA). The obtained bands were quantified in ImageJ x64 by normalizing to loading control and calculating band density relative to untreated control. Resulting graphs show an average of three independent donors.
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