cy5 secondary antibody  (Jackson Immuno)

Journal: Advanced Science

Article Title: A Biomimetic cRGDyk‐Engineered Platelet Nanoplatform for ACAD11 Delivery to Trigger Pyroptosis in Hepatocellular Carcinoma

doi: 10.1002/advs.78219

Figure Lengend Snippet: The construction and characterization of OE‐ACAD11@R‐PLT. (A) Schematic representation of the construction of OE‐ACAD11@R‐PLT. (B) The size distribution of PLT, OE‐ACAD11, cRGDyk, and OE‐ACAD11@R‐PLT. (C) The zeta potential of PLT, cRGDyk, OE‐ACAD11, and OE‐ACAD11@R‐PLT. (D) SEM images of raw PLT and OE‐ACAD11@R‐PLT. (E) The marker protein (CD62P, CD61, CD41, and GAPDH) in raw PLT and OE‐ACAD11@R‐PLT. (F) CLSM results of OE‐ACAD11@R‐PLT. Green, FITC‐labeled PLTs. Red, Cy5‐labeled cRGDyk. (G) Drug loading efficiency and encapsulation rate of OE‐ACAD11@R‐PLT.

Article Snippet: First, the cRGDyk peptide was labeled with CY5 (MCE, USA) via amino‐carboxyl coupling.

Techniques: Zeta Potential Analyzer, Marker, Labeling, Encapsulation

Journal: Bioactive Materials

Article Title: Countering postoperative immune suppression with a self-assembling dendritic cell nanovaccine

doi: 10.1016/j.bioactmat.2026.05.005

Figure Lengend Snippet: BAITs enhance antigen uptake and digestion by DCs under TGFβ stimulation. (A) Schematic illustration of real-time DC migration assay. BAITs in the lower chamber create a chemokine gradient to attract DCs from the upper chamber. (B) Kinetics of DC migration measured as cell index (n = 3; n.s. is P > 0.05, ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (C) Crystal violet staining of non-migrated DCs (upper chamber) and Calcein-AM staining of migrated DCs (lower chamber) showing enhanced DC recruitment by BAITs. (D) The antigen uptake efficiency of DCs under varying antigen concentrations with or without TGFβ (n = 5; ∗∗∗ is P < 0.001, ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (E) Heatmap of immune-related genes in DCs showing upregulation of antigen presentation and co-stimulation markers and downregulation of suppressive factors as antigen concentration increases. (F) Antigen digestion capacity of DCs assessed in the presence or absence of TGFβ (n = 8; ∗∗∗∗ is P < 0.0001 by two-tailed Student's t -test). (G) Fluorescent images showing enhanced antigen uptake in DCs mediated by BAITs (quantified on right; n = 10; ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (H) Flow cytometry analysis of antigen-positive (Cy3 + ) DCs after incubation with free antigen or BAIT (quantified on right; n = 4; ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (I) BAITs improve antigen digestion and uptake of DCs under TGFβ stimulation. DCs were first incubated with Cy5-antigens for 12 h, followed by medium replacement and subsequent treatment with Cy3-antigens or Cy3-BAITs for an additional 6 h. Data are presented as mean ± SD.

Article Snippet: For MHC I analysis, the cells were incubated with rat CoraLite Plus 488 anti-mouse LAMP1 antibody (1:200) and rabbit anti-mouse MHC I antibody (1:100) overnight at 4 °C, followed by Cy5-conjugated anti-rabbit secondary antibody (1:200) for 1 h at 37 °C.

Techniques: Migration, Staining, Immunopeptidomics, Concentration Assay, Two Tailed Test, Flow Cytometry, Incubation