unconjugated anti cd99 antibodies  (Proteintech)

Journal: PLOS One

Article Title: CD99-targeted immunomagnetic negative selection: A novel strategy for high-purity pancreatic islet isolation in murine models

doi: 10.1371/journal.pone.0344446

Figure Lengend Snippet: (A) Immunofluorescence co-localization analysis. Paraffin-embedded mouse pancreatic sections were immunostained with antibodies against insulin (green) and CD99 (red), followed by DAPI counterstaining (blue) for nuclear visualization. CD99 immunoreactivity (red) was exclusively localized to exocrine tissue and did not co-localize with insulin-positive β-cells (green), magnification, 20 × ; scale bar, 100 μm. (B) Immunohistochemical analysis. CD99 expression (brown) was evaluated in paraffin-embedded sections of intact mouse pancreas, purified islets, and isolated exocrine fragments. Robust CD99 immunoreactivity was observed in exocrine regions of intact pancreatic tissue, while islets remained consistently negative. Purified islets exhibited no detectable CD99 expression, magnification, 40 × ; scale bars, 50 μm (pancreas sections), 10 μm (purified islets and exocrine fragments). ( C ) western blot analysis. CD99 protein expression was quantified in purified islets and isolated exocrine fragments. CD99 was selectively detected in exocrine samples while remaining undetectable in islet preparations. β-Actin served as the loading control.

Article Snippet: Biotinylated and unconjugated anti-CD99 antibodies were purchased from Beijing Solarbio Science & Technology Co., Ltd. Anti-insulin antibodies, enhanced chemiluminescence (ECL) reagents, and fluorophore-conjugated secondary antibodies were acquired from Proteintech.

Techniques: Immunofluorescence, Immunohistochemical staining, Expressing, Purification, Isolation, Western Blot, Control

Journal: PLOS One

Article Title: CD99-targeted immunomagnetic negative selection: A novel strategy for high-purity pancreatic islet isolation in murine models

doi: 10.1371/journal.pone.0344446

Figure Lengend Snippet: (A, B) Representative images of dithizone-stained pancreatic digest samples. (A) Unpurified sample demonstrating mixed cellular composition (islets stained red, exocrine tissue grayish-white). (B) Sample after CD99-targeted immunomagnetic purification, showing substantial enrichment of islets.(C, D) Representative images of dithizone-stained samples processed by Ficoll density gradient centrifugation. (C) Initial unpurified digest. (D) Sample after Ficoll purification.(E) Quantitative analysis of islet purity. Initial purity of digests was comparable between groups. CD99-targeted purification achieved a final purity of 93.0 ± 1.4%, significantly higher than the 69.2 ± 1.1% achieved by Ficoll purification. Data represent mean ± SEM; n = 3 per group. **** p < 0.0001.(F) Quantitative analysis of islet recovery rate. The recovery rate after CD99-targeted purification (81.7 ± 2.2%) was significantly higher than that after Ficoll purification (31 ± 1.5%). Data represent mean ± SEM; n = 3 per group. **** p < 0.0001.

Article Snippet: Biotinylated and unconjugated anti-CD99 antibodies were purchased from Beijing Solarbio Science & Technology Co., Ltd. Anti-insulin antibodies, enhanced chemiluminescence (ECL) reagents, and fluorophore-conjugated secondary antibodies were acquired from Proteintech.

Techniques: Staining, Purification, Gradient Centrifugation

Journal: PLOS One

Article Title: CD99-targeted immunomagnetic negative selection: A novel strategy for high-purity pancreatic islet isolation in murine models

doi: 10.1371/journal.pone.0344446

Figure Lengend Snippet: Pancreatic islets (400 IEQ) purified via CD99-based immunomagnetic separation or Ficoll density gradient centrifugation were transplanted beneath the renal capsule of streptozotocin (STZ)-induced diabetic mice. Glycemic control and body weight were monitored over 40 days post-transplantation. (A) Glycemic response. All recipients exhibited hyperglycemia prior to transplantation (Day 0). Both treatment groups demonstrated rapid glucose normalization, achieving euglycemia (<200 mg/dL) by day 3 post-transplantation, which was sustained throughout the observation period. Area under the curve (AUC) analysis revealed no significant difference in overall glycemic control between purification methods, confirming equivalent therapeutic efficacy. n = 5; ns, not significant, p > 0.05. (B) Body weight dynamics. Both groups exhibited initial transient weight loss followed by progressive recovery, consistent with metabolic restoration. No significant inter-group differences were observed throughout the study period. n = 5; ns, not significant, p > 0.05.

Article Snippet: Biotinylated and unconjugated anti-CD99 antibodies were purchased from Beijing Solarbio Science & Technology Co., Ltd. Anti-insulin antibodies, enhanced chemiluminescence (ECL) reagents, and fluorophore-conjugated secondary antibodies were acquired from Proteintech.

Techniques: Purification, Immunomagnetic Separation, Gradient Centrifugation, Control, Transplantation Assay, Drug discovery