mouse anti s100p monoclonal antibodies  (R&D Systems)

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Down-regulation of S100P induces apoptosis in endometrial epithelial cell during GnRH antagonist protocol

doi: 10.1186/s12958-021-00787-0

Figure Lengend Snippet: S100P is down-regulated in the mid-secretory phase endometrium of participants from GnRH-ant group. A S100P gene expression in different groups from microarray data. B Immunohistochemical detection of S100P in the endometrium from the different groups, magnified in 200 times and the scale bar represents 50um. a. Negative control, b. Natural cycle control, c. GnRH antagonist group, d. GnRH agonist group; quantified in ( C ). D Location of S100P in endometrium by co-localization with vimentin and keratin. Red: S100P; Green: vimentin (b) and cytokeratin (f); Blue: nucleus. The scale bar represents 50um. E Western blot results of S100P and HOXA10 protein expression in endometrium from the natural cycle, GnRH-ant and GnRH-a groups. F Quantification of S100P protein expression. G Quantification of HOXA10 protein expression. * P < .05, ** P <.01, *** P <.001

Article Snippet: Briefly, the primary antibodies used are as follows: mouse anti-S100P monoclonal antibodies (MAB2957, R&D Systems, Minneapolis, MN, USA, 1:50 dilution), rabbit anti-cytokeratin monoclonal antibodies (ab181598, Abcam, UK, 1:100), rabbit anti-Vimentin monoclonal antibodies (ab92547, Abcam, UK, 1:100) and isotype antibody as negative control.

Techniques: Gene Expression, Microarray, Immunohistochemical staining, Negative Control, Control, Western Blot, Expressing

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Down-regulation of S100P induces apoptosis in endometrial epithelial cell during GnRH antagonist protocol

doi: 10.1186/s12958-021-00787-0

Figure Lengend Snippet: Knockdown of S100P induces apoptosis in endometrial epithelial cells. A Overexpression and knockdown of S100P mRNA was verified by Realtime PCR. B Overexpression and knockdown of S100P protein was verified and expression of Bcl-2 and Bax protein was analyzed by western blot. C Apoptosis rate was evaluated by flow cytometry after overexpression and knockdown of S100P in Ishikawa cells; results are quantified and analyzed in ( D ). NC: negative control using scramble sequence; KD: S100P knockdown; Mock: mock control transfected with empty plasmid vector; OE: S100P overexpression; * P < .05

Article Snippet: Briefly, the primary antibodies used are as follows: mouse anti-S100P monoclonal antibodies (MAB2957, R&D Systems, Minneapolis, MN, USA, 1:50 dilution), rabbit anti-cytokeratin monoclonal antibodies (ab181598, Abcam, UK, 1:100), rabbit anti-Vimentin monoclonal antibodies (ab92547, Abcam, UK, 1:100) and isotype antibody as negative control.

Techniques: Knockdown, Over Expression, Expressing, Western Blot, Flow Cytometry, Negative Control, Sequencing, Control, Transfection, Plasmid Preparation

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Down-regulation of S100P induces apoptosis in endometrial epithelial cell during GnRH antagonist protocol

doi: 10.1186/s12958-021-00787-0

Figure Lengend Snippet: GnRH antagonist-induced apoptosis in endometrial epithelial cells is rescued by S100P overexpression. A S100P concentration in the cell culture supernatant after GnRH-ant treatment, as determined by ELISA. B Protein expression of S100P, Bcl-2 and Bax in cultured cells after GnRH-ant treatment, as determined by western blot. C Flow cytometry analysis of apoptosis rate among different treatment groups; quantified and analyzed in ( D ). Mock: mock control transfected with empty plasmid vector; ANT: cells exposed to the GnRH antagonist- Cetrorelix; OE: S100P overexpression; * P < .05

Article Snippet: Briefly, the primary antibodies used are as follows: mouse anti-S100P monoclonal antibodies (MAB2957, R&D Systems, Minneapolis, MN, USA, 1:50 dilution), rabbit anti-cytokeratin monoclonal antibodies (ab181598, Abcam, UK, 1:100), rabbit anti-Vimentin monoclonal antibodies (ab92547, Abcam, UK, 1:100) and isotype antibody as negative control.

Techniques: Over Expression, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Flow Cytometry, Control, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Sequences of primers used in the present study

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques:

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Expression of S100P in endometrial cells. A: Immunofluorescence staining for S100P (red) and Hoechst33258 (blue) in EECs (a, b, c), ESCs (d, e, f), Ishikawa cells (g, h, i), and RL95–2 cells (j, k, l). PBS was used as a negative control (m, n, o), and the scale bar was 50 μm). B: The mRNA expression of S100P in different endometrial cells, as determined using qPCR. C: Fluorescence densitometric analysis using image J, normalized to that of EECs. EEC: primary endometrial epithelial cells, ESC: primary endometrial stromal cells, * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques: Expressing, Immunofluorescence, Staining, Negative Control, Fluorescence

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Expression of S100P in endometrial cancer and its precursor lesions. A: Immunohistochemistry demonstrating the distribution of S100P in endometrial cancer and its precursor lesions. a, Negative control; b, Normal endometrium in the proliferative phase; c, Endometrium of SH; d, Endometrium of CH; e, Endometrium of AH; f, Endometrium of EC (Magnified 200×). B: Densitometric analysis using image J. The data were normalized to the normal control. ns: no significance; * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques: Expressing, Immunohistochemistry, Negative Control, Control

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: The efficiency of S100P knockdown and overexpression in cells. A, C: Knockdown of S100P (red) in Ishikawa or RL95–2 cells, respectively; Hoechst33258 (blue) for nuclear staining (Magnified 200 times). B, D: The changes in the S100P mRNA level in A or C, as determined using qPCR. ACTB (β-actin) was used as the internal reference (* P < 0.05). E, F: overexpression of S100P (red) in primary endometrial stromal cells and changes in the S100P mRNA level as mentioned above. Negative: uninfected cells; S100P shRNA: cells infected with S100P interference lentiviruses; S100P overexpression: cells infected with S100P overexpression lentiviruses; Control: Cells infected with scrambled sequence lentiviruses (A, B, C, D) or empty vector (E, F)

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques: Knockdown, Over Expression, Staining, shRNA, Infection, Control, Sequencing, Plasmid Preparation

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Transwell experiments showing the effects of S100P on cell invasion. A: The invasive capacity of Ishikawa cells or endometrial stromal cells after silencing or overexpression of S100P using lentiviruses was determined using Transwell assays. Representative images (magnified 200 times) were captured at 48 h after the cells were seeded. All the experiments were performed in triplicate. a, b: ESCs transfected with the empty vector or showing S100P overexpression, respectively; c, d: Ishikawa cells with the scramble vector or subjected to S100P downregulation, respectively. B and C: Histogram showing the number of invasive cells per field from three independent experiments. * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques: Over Expression, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: The colocalization of S100P and Ezrin, and F-actin rearrangement after knocking down S100P . A: Laser confocal microscopy showing the colocalization (yellow) of S100P (green) and Ezrin (red) in RL95–2 cells. The panels below (e, f, g, h) are an enlargement of the box area (a, b, c, d). B and C: FITC-phalloidin (green) staining showing the rearrangement of F-actin in RL95–2 cells (left) and Ishikawa cells (right). Scramble control (a, c, e) or S100P shRNA (b, d, f) set as mentioned previously. The nuclei were counterstained using Hoechst33258 (blue). The magnification of (A) is displayed using a ruler and the magnification in B and C was 400 ×

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, cells were incubated with 1:50 mouse anti-human S100P monoclonal antibody (R&D Systems, Minneapolis, MN, USA) as the primary antibody, and then incubated with 1:500 Cy3-labeled goat anti-mouse IgG (Invitrogen) as the secondary antibody.

Techniques: Confocal Microscopy, Staining, Control, shRNA

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Sequences of primers used in the present study

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques:

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Expression of S100P in endometrial cells. A: Immunofluorescence staining for S100P (red) and Hoechst33258 (blue) in EECs (a, b, c), ESCs (d, e, f), Ishikawa cells (g, h, i), and RL95–2 cells (j, k, l). PBS was used as a negative control (m, n, o), and the scale bar was 50 μm). B: The mRNA expression of S100P in different endometrial cells, as determined using qPCR. C: Fluorescence densitometric analysis using image J, normalized to that of EECs. EEC: primary endometrial epithelial cells, ESC: primary endometrial stromal cells, * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques: Expressing, Immunofluorescence, Staining, Negative Control, Fluorescence

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Expression of S100P in endometrial cancer and its precursor lesions. A: Immunohistochemistry demonstrating the distribution of S100P in endometrial cancer and its precursor lesions. a, Negative control; b, Normal endometrium in the proliferative phase; c, Endometrium of SH; d, Endometrium of CH; e, Endometrium of AH; f, Endometrium of EC (Magnified 200×). B: Densitometric analysis using image J. The data were normalized to the normal control. ns: no significance; * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques: Expressing, Immunohistochemistry, Negative Control, Control

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: The efficiency of S100P knockdown and overexpression in cells. A, C: Knockdown of S100P (red) in Ishikawa or RL95–2 cells, respectively; Hoechst33258 (blue) for nuclear staining (Magnified 200 times). B, D: The changes in the S100P mRNA level in A or C, as determined using qPCR. ACTB (β-actin) was used as the internal reference (* P < 0.05). E, F: overexpression of S100P (red) in primary endometrial stromal cells and changes in the S100P mRNA level as mentioned above. Negative: uninfected cells; S100P shRNA: cells infected with S100P interference lentiviruses; S100P overexpression: cells infected with S100P overexpression lentiviruses; Control: Cells infected with scrambled sequence lentiviruses (A, B, C, D) or empty vector (E, F)

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques: Knockdown, Over Expression, Staining, shRNA, Infection, Control, Sequencing, Plasmid Preparation

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: Transwell experiments showing the effects of S100P on cell invasion. A: The invasive capacity of Ishikawa cells or endometrial stromal cells after silencing or overexpression of S100P using lentiviruses was determined using Transwell assays. Representative images (magnified 200 times) were captured at 48 h after the cells were seeded. All the experiments were performed in triplicate. a, b: ESCs transfected with the empty vector or showing S100P overexpression, respectively; c, d: Ishikawa cells with the scramble vector or subjected to S100P downregulation, respectively. B and C: Histogram showing the number of invasive cells per field from three independent experiments. * P < 0.05

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques: Over Expression, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Promotion of the occurrence of endometrioid carcinoma by S100 calcium binding protein P

doi: 10.1186/s12885-020-07350-x

Figure Lengend Snippet: The colocalization of S100P and Ezrin, and F-actin rearrangement after knocking down S100P . A: Laser confocal microscopy showing the colocalization (yellow) of S100P (green) and Ezrin (red) in RL95–2 cells. The panels below (e, f, g, h) are an enlargement of the box area (a, b, c, d). B and C: FITC-phalloidin (green) staining showing the rearrangement of F-actin in RL95–2 cells (left) and Ishikawa cells (right). Scramble control (a, c, e) or S100P shRNA (b, d, f) set as mentioned previously. The nuclei were counterstained using Hoechst33258 (blue). The magnification of (A) is displayed using a ruler and the magnification in B and C was 400 ×

Article Snippet: After permeabilization with 0.1% Triton X-100 for 45 min, rabbit anti-human Ezrin 1: 50 dilution (Epitomics, Burlingame, CA, USA; EP886Y) and mouse anti-human S100P 1:50 dilution (R&D Systems) were used as the primary antibodies.

Techniques: Confocal Microscopy, Staining, Control, shRNA

Journal: BioMed Research International

Article Title: Differentiation Model Establishment and Differentiation-Related Protein Screening in Primary Cultured Human Sebocytes

doi: 10.1155/2018/7174561

Figure Lengend Snippet: Skin-associated functions of 41 differentially expressed proteins at D7 compared with D0.

Article Snippet: The primary antibodies included polyclonal mouse anti-human S100P (clone B10, 1 : 100 dilution; Santa Cruz), ADA (clone D10, 1 : 100 dilution; Santa Cruz), K10 (clone LH1, 1 : 100 dilution; Santa Cruz), and rabbit anti-human FDXR (ab122900, 1 : 100 dilution; Abcam).

Techniques: Migration, Binding Assay, Activation Assay, Infection, Ubiquitin Proteomics, Expressing, Cell Differentiation, Membrane

Journal: BioMed Research International

Article Title: Differentiation Model Establishment and Differentiation-Related Protein Screening in Primary Cultured Human Sebocytes

doi: 10.1155/2018/7174561

Figure Lengend Snippet: Expression of 4 candidate proteins in sebocytes and acne lesion. (a) Human primary sebocytes were cultivated under serum deprivation for indicated times, and expression levels of S100P, FDXR, ADA, and K10 were monitored by Western blot. Actin was used as a loading control. (b) Representative immunostaining images of S100P, FDXR, ADA, and K10 in sebaceous glands of acne lesion and normal skin (scale bars = 50 μ m).

Article Snippet: The primary antibodies included polyclonal mouse anti-human S100P (clone B10, 1 : 100 dilution; Santa Cruz), ADA (clone D10, 1 : 100 dilution; Santa Cruz), K10 (clone LH1, 1 : 100 dilution; Santa Cruz), and rabbit anti-human FDXR (ab122900, 1 : 100 dilution; Abcam).

Techniques: Expressing, Western Blot, Control, Immunostaining