
Western blot analysis of lysates from wild type (WT) and Annexin A1/ANXA1 knockdown (KD) HeLa cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at 1:50000 dilution incubated at room temperature for 1.5 hours.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).Exposure time: 1s.

Western blot analysis of various lysates using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at 1:50000 dilution incubated at room temperature for 1.5 hours.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).Exposure time: 1s.

Immunohistochemistry analysis of paraffin-embedded Human liver cancer tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Hela and HeLa-ANXA1-KO cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Confocal imaging of HeLa cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of C2C12 cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of U-251 MG cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Immunoprecipitation of [KD Validated] Annexin A1/ANXA1 from 500 µg extracts of C2C12 cells was performed using 2 µg of [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:8000.

Flow cytometry: 1X10^6 293T cells (negative control,left) and A549 cells (right) were intracellularly-stained with [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:1000,orange line) or Rabbit IgG isotype control (AC042,2 μg/mL,blue line), followed by APC conjugated goat anti-rabbit pAb staining. Non-fluorescently stained cells were used as blank control (red line).

Flow cytometry: 1X10^6 A549 cells (right) were intracellularly-stained with Rabbit IgG isotype control (AC042,2 μg/mL,left) or [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:1000,right), followed by APC conjugated goat anti-rabbit pAb staining.

Flow cytometry: 1X10^6 Human PBMC (right) were intracellularly-stained with Rabbit IgG isotype control (AC042,2 μg/mL,left) or [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:1000,right), followed by APC conjugated goat anti-rabbit pAb staining. Cells in the monocyte gate were used for analysis.