Syk Rabbit mAb (A25741)

Western blot analysis of various lysates using Syk Rabbit mAb (A25741)at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020)
Negative control (NC): MOLT-4
Exposure time: 45 s.

Confocal imaging of Daudi cells using Syk Rabbit mAb (A25741, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of HEL cells using Syk Rabbit mAb (A25741, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of A20 cells using Syk Rabbit mAb (A25741, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of paraffin-embedded Rat spleen tissue using Syk Rabbit mAb (A25741, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.

Immunoprecipitation of Syk from 500 µg extracts of HEL cells was performed using 2 µg of Syk Rabbit mAb (A25741). Rabbit IgG isotype control(AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X non-reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using Syk Rabbit mAb (A25741) at a dilution of 1:1000.

Flow cytometry: 1X10^6 Human PBMC were surface-stained with FITC Mouse anti-Human CD19 mAb (A22815, 5 μl/Test) followed by intracellular staining with Syk Rabbit mAb (A25741, 5 μl/Test, left) or Rabbit IgG isotype control (AC042, 2 μg/mL, right).