Anti-p19 INK4d antibody [RP23040113]

• IP

Supplier Data

Immunoprecipitation - Anti-p19 INK4d antibody [RP23040113] (AB308000)

P19INK4D was immunoprecipitated using 2 µg of ab30800 from lysate of HeLa cells treated with Trichostatin A (2 uM/24 hours) (Lane 3). Normal Rabbit IgG was used as a negative IP control (Lane 2). 10% input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using ab307800 and Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 2500 dilution. Chemiluminescent detection was performed (ECL).

All lanes:

Immunoprecipitation - Anti-p19 INK4d antibody [RP23040113] (ab308000) at 2 µg

Lane 1:

HeLa cell lysate

Lane 2:

HeLa cell lysate with normal rabbit IgG

Lane 3:

HeLa cells lysate treated with Trichostatin A

Secondary

All lanes:

HRP-conjugated Goat anti-Rabbit IgG (H+L) Secondary Antibody at 1/2500 dilution

Observed band size: 18 kDa

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• WB

Supplier Data

Western blot - Anti-p19 INK4d antibody [RP23040113] (AB308000)

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK- 293 (Lane1), HEK- 293 treated with Trichostatin A, (Lane 2), HeLa (Lane 3), HeLa treated with Trichostatin A (Lane 4), HCT 116 (Lane 5), HCT 116 treated with Trichostatin A (Lane 6) (2µM/24hrs). The blots were probed with ab308000 at 0.5-1 µg/mL and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 2500 dilution. A clear 18 kDa band corresponding to P19INK4D was observed across cell lines tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate

All lanes:

Western blot - Anti-p19 INK4d antibody [RP23040113] (ab308000) at 0.5 µg/mL

Lane 1:

Untreated HEK- 293 at 30 µg

Lane 2:

HEK- 293 treated with Trichostatin A at 30 µg

Lane 3:

Untreated HeLa at 30 µg

Lane 4:

HeLa treated with Trichostatin A at 30 µg

Lane 5:

HCT 116 at 30 µg

Lane 6:

HCT 116 treated with Trichostatin A at 30 µg

Secondary

All lanes:

HRP-conjugated Goat anti-Rabbit IgG (H+L) Secondary Antibody at 1/2500 dilution

Observed band size: 18 kDa

true