Anti-CacyBP antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CacyBP antibody (AB190950)

CACYBP was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL ab190950 overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1 : 500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI.

• Flow Cyt

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Flow Cytometry - Anti-CacyBP antibody (AB190950)

Overlay histogram showing HL-60 cells stained with ab190950 (Blue). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with ab190950 1 µg/1x10^6 cells for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG 5-10 µg/1x10^6 cells was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red) was also used as a control.

• IP

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Immunoprecipitation - Anti-CacyBP antibody (AB190950)

CacyBP was immunoprecipitated from U251 whole cell lysate with ab190950. Western blot was performed on the immunoprecipitate using ab190950 at 0.5 ug/ml followed by a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate.

Lane 1 : U251 whole cell lysates, 30 ug
Lane 2 : Rabbit control IgG instead of anti-CACYBP antibody in U251 whole cell lysate
Lane 3 : Ranti-CACYBP antibody (2μg) + U251 whole cell lysate (500μg)

Lanes 1 and 3:

Immunoprecipitation - Anti-CacyBP antibody (ab190950) at 0.5 µg/mL

Lane 2:

Rabbit control IgG

Lane 1:

U251 whole cell lysates at 30 µg

Lane 2:

Rabbit control IgG instead of anti-CACYBP antibody in U251 whole cell lysate

Lane 3:

Ranti-CACYBP antibody (2μg) + U251 whole cell lysate (500μg)

Secondary

All lanes:

goat anti-rabbit IgG-HRP secondary antibody

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

• WB

Supplier Data

Western blot - Anti-CacyBP antibody (AB190950)

Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with ab190950 at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1/5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CACYBP at approximately 27 kDa. The expected band size for CACYBP is at 27 kDa.

All lanes:

Western blot - Anti-CacyBP antibody (ab190950) at 0.5 µg/mL

Lane 1:

human MCF-7 whole cell lysates, at 30 µg

Lane 2:

human RT4 whole cell lysates, at 30 µg

Lane 3:

human SW620 whole cell lysates, at 30 µg

Lane 4:

human U251 whole cell lysates, at 30 µg

Lane 5:

rat liver tissue lysates, at 30 µg

Lane 6:

rat brain tissue lysates, at 30 µg

Lane 7:

mouse liver tissue lysates, at 30 µg

Lane 8:

mouse brain tissue lysates. at 30 µg

Secondary

All lanes:

goat anti-rabbit IgG-HRP at 1/5000 dilution

true