- PA5-28988 - Invitrogen Antibodies CSF3R antibody

Ognibene M, Pezzolo A
Scientific reports 2020 Jul 31;10(1):12902

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of G-CSF Receptor/CD114 using 30 µg THP-1 whole cell lysate. Samples were loaded onto a 5% SDS-PAGE gel and probed with a G-CSF Receptor/CD114 polyclonal antibody (Product # PA5-28988) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of G-CSF Receptor/CD114 using Whole cell extracts (30 µg). Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a G-CSF Receptor/CD114 polyclonal antibody (Product # PA5-28988) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of CSF3R was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a CSF3R Polyclonal Antibody (Product # PA5-28988) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: CSF3R Polyclonal Antibody detects CSF3R protein at cytoplasm by immunofluorescent analysis. Sample: THP-1 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: CSF3R protein stained by CSF3R Polyclonal Antibody (Product # PA5-28988) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: CSF3R Polyclonal Antibody detects GCSF Receptor protein at secreted on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. CSF3R Polyclonal Antibody (Product # PA5-28988) diluted at 1:250. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Neuroblastoma tumor spheres show high levels of cancer stem cell surface markers and of key nucleolar proteins. ( A ) Neurospheres derived from IMR-32, ACN and SH-SY5Y neuroblastoma cell lines cultured for three days in serum-free medium and in non-adherent conditions. Spheres show differences in number and dimensions (left photos), cells aggregation (middle photos) and adhesion ability (right photos). (Scale bars: 200 um on the left and 100 um in the middle and on the right). ( B ) Protein lysates from IMR-32, ACN and SH-SY5Y cell lines and from neurospheres derived by each cell line were collected and subjected to Western blot analysis with anti-CD44v6, anti-CD114, anti-Nucleolin (NCL), anti-Nucleophosmin-1 (NPM1), anti-Glypican-2 (GPC2) and anti-Pescadillo Ribosomal Biogenesis Factor-1 (PES1) antibodies. Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Supplementary Figure S5 shows the entire blots images. ( C ) Neurospheres protein levels were quantified by densitometry, normalized to those of each cell line (fold induction = 1) and to the content of the loading control protein (Actin), then visualized by histograms. Data are representative of three independent experiments +- SD (*** p < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Roniciclib strongly inhibits cancer stem cells markers expression while enhances p53 onco-suppressor level in neuroblastoma cells. ( A ) Protein lysates from IMR-32, ACN and SH-SY5Y cell lines untreated (u = Roniciclib 0 uM) or treated with Roniciclib (R) 1 uM, 20 uM and 5 uM respectively, for 72 h were collected and subjected to Western blot analysis with anti-CD44v6, anti-CD114, anti-Osteopontin (OPN), anti-Microtubule-associated protein-2 (MAP2), anti-p53, anti-beta-catenin and anti-Low density lipoprotein related protein (LRP6) antibodies. Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Supplementary Figure S6 , panels 1-2, shows the entire blots images. ( B ) Protein levels of the Roniciclib treated cells were quantified by densitometry, normalized to those of the untreated cells (fold induction = 1) and to the content of the loading control protein (Actin), then visualized by histograms. Data are representative of three independent experiments +- SD (* p < 0.05; ** p < 0.01; *** p < 0.001).

PA5-28988