human stc2 neutralization antibodies  (R&D Systems)

Journal: Annals of Medicine and Surgery

Article Title: Inhibiting stanniocalcin 2 reduces sunitinib resistance of Caki-1 renal cancer cells under hypoxia condition

doi: 10.1097/MS9.0000000000001450

Figure Lengend Snippet: Effect of sunitinib on STC2 expression in Caki-1 cells under hypoxia condition. (A) The mRNA level of STC2 in Caki-1 cells was evaluated by RT-qPCR. (B, C) The protein levels of STC2 in Caki-1 cell lines were evaluated by Western blotting after treated with hypoxia mimic CoCl 2 (100 μM) or treated in hypoxic incubator for 24 h. (D, E) The protein levels of STC2 in Caki-1 cells were evaluated by Western blotting after treated with or without Sun (5 μM). Relative protein levels were normalized by β-Actin and analyzed by Image J 1.54c. *, P <0.05 indicates significant difference. Sun, sunitinib.

Article Snippet: Human STC2-neutralization antibodies (AF2830) and recombinant human STC2 (O76061) were purchased from R&D System (USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Annals of Medicine and Surgery

Article Title: Inhibiting stanniocalcin 2 reduces sunitinib resistance of Caki-1 renal cancer cells under hypoxia condition

doi: 10.1097/MS9.0000000000001450

Figure Lengend Snippet: Regulation of STC2 on sunitinib efficacy in Caki-1 cells under hypoxia condition. Determination of cell viability of Caki-1 cells treated with or without 5 μM Sun, 5 μM sunitinib + 0.2 μg/ml STC2-neutralizing antibodies siRNA-STC2 knockdown + 5 μM Sun or 5 μM Sun + 500 ng/ml hSTC2 by MTT (A). The cell proliferation of Caki-1 cells was determined by immunofluorescence staining of proliferation maker Ki-67 and quantified by the software Image J 1.54c (B, C). The cells were incubated with 5 μM Sun + 0.2 μg/ml STC2-neutralizing antibodies, or 5 μM Sun + 500 ng/ml hSTC2 for 24 h, the accumulation of s Sun in Caki-1 cells were observed by phase contrast microscopy (D). * P <0.05, ** P <0.01, indicate significant difference. Sun, sunitinib.

Article Snippet: Human STC2-neutralization antibodies (AF2830) and recombinant human STC2 (O76061) were purchased from R&D System (USA).

Techniques: Knockdown, Immunofluorescence, Staining, Software, Incubation, Microscopy

Journal: Annals of Medicine and Surgery

Article Title: Inhibiting stanniocalcin 2 reduces sunitinib resistance of Caki-1 renal cancer cells under hypoxia condition

doi: 10.1097/MS9.0000000000001450

Figure Lengend Snippet: Effects of STC2-neutralizing antibodies on EMT, migration and invasion of Caki-1 cells treated with sunitinib under hypoxia condition. After treated with or without 5 μM Sun, 5 μM Sun + 0.2 μg/ml STC2-neutralizing antibodies for 24 h, the protein levels of EMT biomarkers in Caki-1 cells were evaluated by Western blotting (A), and quantified by Image J (B). The cell migration and invasion of Caki-1 cells were evaluated by wound healing assay and transwell (C-F). *, P <0.05; **, P <0.01 indicate significant difference. Sun, sunitinib.

Article Snippet: Human STC2-neutralization antibodies (AF2830) and recombinant human STC2 (O76061) were purchased from R&D System (USA).

Techniques: Migration, Western Blot, Wound Healing Assay

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 1. Expression pattern of STC2 in KIRC specimens and ccRCC cell lines. A) GEPIA analysis shows the mRNA expression levels of STC2 in 523 KIRCs and 100 non-KIRCs. B, C) The relative expression levels of STC2 in HK-2 cells and ccRCC cell lines were evalu ated by RT-qPCR and western blotting. β-actin was used as a loading reference. D) The STC2 contents in the culture medium of HK-2 cells and ccRCC cell lines were detected by ELISA. *p<0.05, **p<0.01

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 3. Effect of STC2 on sunitinib resistance in ccRCC cells. A–C) Determi nation of cell viability of Caki-1, 786-O, and 769-P cells in the absence (Ctrl) or presence of 5 μM sunitinib or 5 μM sunitinib+0.2 μg/ml STC2 neutralizing an tibody. D–F) Determination of cell viability of Caki-1, 786-O, and 769-P cells in the absence (Ctrl) or presence of 5 μM sunitinib or 5 μM sunitinib+500 ng/ ml hSTC2. *p<0.05

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques:

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 6. Effects of the STC2 neutralizing antibody on sunitinib accumulation, lysosomal pH, cell proliferation, and apoptosis in Caki-1 cells. A) Phase-contrast microscopy showing accumulation of yellow granules in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. B, C) AO staining indicates the lysosome pH in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. Quantification of the ratio between the red and green signal of AO was performed by the software ImageJ. D) The fluorescence of the lysosomal probe (LysoTracker™ Green DND-26) in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. E, F) The immuno fluorescence staining of cell proliferation marker Ki-67 and cell apoptosis detection by TUNEL staining in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. Quantification of Ki-67 and TUNEL fluorescence was performed by the software ImageJ. NS: no significant difference between control with treatments. *p<0.05

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques: Microscopy, Incubation, Staining, Software, Fluorescence, Marker, TUNEL Assay, Control