rabbit polyclonal anti human igg fc  (Bethyl)

Journal: Scientific reports

Article Title: Altered sialidase expression in human myeloid cells undergoing apoptosis and differentiation.

doi: 10.1038/s41598-022-18448-6

Figure Lengend Snippet: Figure 4. NEU and PPCA Gene Expression During HL60 Cell Differentiation. (A) Total RNA isolated from nondifferentiated HL60 cells was processed for qRT-PCR for mRNA levels for NEU1-4 and PPCA, and normalized to the 18S rRNA control. (B–F) Total RNA isolated from HL60 cells cultured for increasing times in the presence of DMF or medium alone was processed for qRT-PCR to quantify mRNA levels for NEU1 (B), PPCA (C), NEU2 (D), NEU3 (E), and NEU4 (F), and each normalized to the 18S rRNA control. The n for each data point is indicated below each vertical bar (A) or time point (B–F). Each vertical bar (A) or symbol (B–F) represents mean ± SE normalized mRNA levels. The data generated in each panel represents experiments performed on ≥ 2 independent occasions. *, increased normalized NEU2/4 mRNA expression compared to nondifferentiated cells at p < 0.05.

Article Snippet: In some experiments, the blots were blocked for 1 h using 5.0% nonfat milk in 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, and 0.01% Tween 20 (TBS-T), probed with murine anti-human NEU1 monoclonal antibody (OriGene, Rockville, MD)45,47, rabbit anti-human PPCA monoclonal antibody (Abcam, Cambridge, MA)45, rabbit anti-human NEU2 polyclonal antibody (Invitrogen, Thermo Fisher Scientific, Rockford, IL)48, rabbit anti-human NEU3 polyclonal antibody (Novus, Centennial, CO)43, or rabbit anti-human NEU4 polyclonal antibody (Invitrogen)48, each followed by either HRP-conjugated goat anti-rabbit antibody or horse anti-mouse antibody (Cell Signaling, Danvers, MA), and developed with enhanced chemiluminescence (ECL) reagents.

Techniques: Gene Expression, Cell Differentiation, Isolation, Quantitative RT-PCR, Control, Cell Culture, Generated, Expressing

Journal: Scientific reports

Article Title: Altered sialidase expression in human myeloid cells undergoing apoptosis and differentiation.

doi: 10.1038/s41598-022-18448-6

Figure Lengend Snippet: Figure 5. NEU and PPCA Protein Expression During HL60 Cell Differentiation. HL60 cells were cultured for 1, 3, 5 or 7 days in the presence of DMF or medium alone, lysed, and the lysates, at 50 µg total cellular protein/lane, were processed for NEU1 (A), PPCA (C), NEU2 (E), NEU3 (G), and NEU4 (I) immunoblotting. (A, C, E, G, and I) to control for protein loading and transfer, blots were stripped and reprobed for β- actin. IB, immunoblot; IB*, immunoblot after stripping. MW in kDa is indicated on the left. (B, D, F, H, and J) densitometric analyses of the blots in A, C, E, G, and I, respectively. Vertical bars represent mean ± SE NEU1, PPCA, NEU2, NEU3, or NEU4 signal normalized to β-actin signal in the same lane on the same stripped and reprobed blot. *, increased normalized PPCA, NEU2, or NEU4 signal in dHL60 cells compared to nondifferentiated cells at p < 0.05. **, decreased normalized NEU1 signal in dHL60 cells compared to nondifferentiated cells at p < 0.05. Each blot is representative of 3–7 independent experiments. Cropped immunoblot images are shown.

Article Snippet: In some experiments, the blots were blocked for 1 h using 5.0% nonfat milk in 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, and 0.01% Tween 20 (TBS-T), probed with murine anti-human NEU1 monoclonal antibody (OriGene, Rockville, MD)45,47, rabbit anti-human PPCA monoclonal antibody (Abcam, Cambridge, MA)45, rabbit anti-human NEU2 polyclonal antibody (Invitrogen, Thermo Fisher Scientific, Rockford, IL)48, rabbit anti-human NEU3 polyclonal antibody (Novus, Centennial, CO)43, or rabbit anti-human NEU4 polyclonal antibody (Invitrogen)48, each followed by either HRP-conjugated goat anti-rabbit antibody or horse anti-mouse antibody (Cell Signaling, Danvers, MA), and developed with enhanced chemiluminescence (ECL) reagents.

Techniques: Expressing, Cell Differentiation, Cell Culture, Western Blot, Control, Stripping Membranes

Journal: Scientific reports

Article Title: Altered sialidase expression in human myeloid cells undergoing apoptosis and differentiation.

doi: 10.1038/s41598-022-18448-6

Figure Lengend Snippet: Figure 6. NEU and PPCA Protein Expression in Response to Multiple Differentiation Agents. HL60 cells were cultured for 7 days in the presence of DMF, DMSO, RA, or medium alone, lysed, and the lysates, at 50 µg total cellular protein/lane, were processed for NEU1 (A), PPCA (C), NEU2 (E), and NEU3 (G) immunoblotting. For the NEU4 immunoblots, cells were harvested on day 3 (I). (A, C, E, G, and I), to control for protein loading and transfer, blots were stripped and reprobed for β-actin. IB, immunoblot; IB*, immunoblot after stripping. MW in kDa is indicated on the left. (B, D, F, H, and J), densitometric analyses of the blots in (A, C, E, G, and I), respectively. n for each experimental group indicated in each panel (B and D) or under each vertical bar (F, H, and J). Vertical bars represent mean ± SE NEU1, PPCA, NEU2, NEU3, or NEU4 signal normalized to β-actin signal in the same lane on the same stripped and reprobed blot. *, increased normalized PPCA, NEU2, or NEU4 signal in dHL60 cells compared to HL60 cells at p < 0.05. **, decreased normalized NEU1 signal in dHL60 cells compared to HL60 cells at p < 0.05. Each blot is representative of 3–7 independent experiments. Cropped immunoblot images are shown.

Article Snippet: In some experiments, the blots were blocked for 1 h using 5.0% nonfat milk in 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, and 0.01% Tween 20 (TBS-T), probed with murine anti-human NEU1 monoclonal antibody (OriGene, Rockville, MD)45,47, rabbit anti-human PPCA monoclonal antibody (Abcam, Cambridge, MA)45, rabbit anti-human NEU2 polyclonal antibody (Invitrogen, Thermo Fisher Scientific, Rockford, IL)48, rabbit anti-human NEU3 polyclonal antibody (Novus, Centennial, CO)43, or rabbit anti-human NEU4 polyclonal antibody (Invitrogen)48, each followed by either HRP-conjugated goat anti-rabbit antibody or horse anti-mouse antibody (Cell Signaling, Danvers, MA), and developed with enhanced chemiluminescence (ECL) reagents.

Techniques: Expressing, Cell Culture, Western Blot, Control, Stripping Membranes

Journal: STAR Protocols

Article Title: Preparation of monovalent follistatin-like 3-Fc-fusion protein and evaluation of its effects on muscle mass in mice

doi: 10.1016/j.xpro.2021.100839

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-human IgG Fc , Bethyl Laboratories , Cat#A80-105A; RRID: AB_67482.

Techniques: Recombinant, Antibody Purification, Expressing, Enzyme-linked Immunosorbent Assay, Software, Cell Counting, Microscopy