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).
Ab28283
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab28283 is a monoclonal antibody that recognizes the human Transferrin Receptor protein. The antibody is intended for use in various research applications.
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)
Ab215434, 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)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Sc 8059, by Santa Cruz Biotechnology (1 mentions)
Ab32087, by Abcam (1 mentions)
Ab13654, by Abcam (1 mentions)
Ab10540, by Abcam (1 mentions)
Ap132p, by Merck Group (1 mentions)
Immobilon western chemiluminescent hrp substrate wbklso500, by Merck Group (1 mentions)
Ap124p, by Merck Group (1 mentions)
M per, by Thermo Fisher Scientific (1 mentions)
P8340, by Merck Group (1 mentions)
A5060, by Merck Group (1 mentions)
5 protocols using ab28283
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
Embryonic Protein Expression Analysis
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Embryos were dissected from pregnant females at gestational stage 8.5~9.5 and the deciduas were torn off. Total proteins were extracted with Mammalian Protein Extraction Reagent (M-PER; Thermo Scientific, Waltham, MA, USA) containing a protease inhibitor cocktail (P8340, Sigma-Aldrich). Protein concentrations were determined with a Pierce BCA protein assay kit (Thermo Scientific). Appropriate amounts of the extracts were fractionated by SDS-PAGE, electrotransferred to PVDF membranes, and detected with antibodies against CUL4B (HPA011880, Sigma-Aldrich), β-catenin (14–6765, ebioscience, San Diego, CA, USA), phospho-β-catenin (S33/S37/T41) (9561, Cell Signaling Technology, Danvers, MA, USA), cyclin D1 (ab10540, Abcam, Cambridge, MA, USA), cyclin D2 (ab3087, Abcam), cyclin D3 (ab28283, Abcam), cyclin E (630701, Biolegend, San Diego, CA, USA), and β-actin (A5060, Sigma-Aldrich). Signals from the reaction with anti-mouse IgG and anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibodies (AP124P and AP132P, Millipore) were developed with the Immobilon Western Chemiluminescent HRP substrate (WBKLSO500, Millipore). The signals were semiquantified using Multi Gauge V3.0 software (Fujfilm) and normalized against that of β-actin.
5
Detecting Fusion Proteins in Osteosarcoma
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The detection of possible fusion proteins in osteosarcoma tissue lysate and FFPE tissue sections by western blotting and immunohistochemistry used anti-CCND3, anti-KCNMB4, anti-LRP-1 antibodies (ab28283, ab89703 and ab92544, ABCAM, Cambridge, MA) and anti-SNRNP25 antibody (H00079622-B01, Novus, Littleton, CO).
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