anti cd22  (R&D Systems)

Journal: bioRxiv

Article Title: Oxidative Stress-Induced Microglial CD22 Upregulation Impairs Phagocytosis and Exacerbates Huntington’s Disease

doi: 10.64898/2026.03.11.710967

Figure Lengend Snippet: The ITAM-bearing Dectin-1 ( Clec7a ) was cloned from C57BL/6 mice and tagged with V5. Clec7a -V5 was overexpressed in HEK293T cells along with either vector control, CD22 WT , or CD22 R130A . ( a ) Proximity ligation assay (PLA) using anti-CD22 and anti-V5 antibodies revealed molecular proximity between Clec7a -V5 and CD22 WT or CD22 R130A . Data are presented as mean ± SEM (n = 3 independent replicates). Overexpression of Clec7a -V5 in HEK293T cells enabled the uptake of pHRodo red Zymosan bioparticles, and co-expression of CD22 WT or CD22 R130A ( b ) significantly suppressed this uptake, while co-expression of CD22 Y783/843/863F ( c ) did not. Data are presented as mean ± SEM (n = 3 independent replicates). ** p < 0.01, *** p < 0.001 and **** p < 0.0001 by one-way ANOVA. Scale bar = 10 µm.

Article Snippet: After blocking with 4% BSA for 1 hour at room temperature, samples were incubated with anti-CD22 (R&D Systems; AF2296) and anti-V5 (Invitrogen; SV5-Pk1) overnight at 4 °C in a humidified chamber.

Techniques: Clone Assay, Plasmid Preparation, Control, Proximity Ligation Assay, Over Expression, Expressing

Journal: bioRxiv

Article Title: Oxidative Stress-Induced Microglial CD22 Upregulation Impairs Phagocytosis and Exacerbates Huntington’s Disease

doi: 10.64898/2026.03.11.710967

Figure Lengend Snippet: BV2-CD22 microglia were treated with either IgG isotype control or anti-CD22 antibody (Cy34.1) for 24 hours. ( a ) Immunofluorescence staining showed colocalization of CD22 (red) and Cy34.1 (grey) puncta (yellow arrows) within the cells. ( b ) Quantification of GFP showed that cells treated with IgG or Cy34.1 had comparable levels of CD22-GFP, while treatment with Cy34.1 reduced the overall amount of CD22. ( c ) Flow cytometry analysis confirmed similar CD22⁺ percentages (∼99.8%) across groups, but treatment with Cy34.1 reduced the amount of CD22 on the cell surface, as reflected by MFI. Data are presented as mean ± SEM (n = 3). ( d ) Phagocytosis of pHRodo red Zymosan particles was restored in BV2-CD22 cells treated with Cy34.1. Data are presented as mean ± SEM (n = 4 independent experiments). ( e-f ) Phagocytosis of pHRodo-labeled myelin debris was assessed in BV2-CD22 cells treated with Cy34.1 and neonatal primary microglia derived from wild-type (CD22 WT ) or CD22-knockout (CD22 KO ) mice. Phagocytic index was quantified as the percentage of pHRodo⁺ among CD11b⁺ microglia. Data are presented as mean ± SEM (n = 4-5 biological replicates). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 by unpaired t -test. Scale bar = 10 µm.

Article Snippet: After blocking with 4% BSA for 1 hour at room temperature, samples were incubated with anti-CD22 (R&D Systems; AF2296) and anti-V5 (Invitrogen; SV5-Pk1) overnight at 4 °C in a humidified chamber.

Techniques: Control, Immunofluorescence, Staining, Flow Cytometry, Labeling, Derivative Assay, Knock-Out