Anti-PUMA antibody

• ICC

Supplier Data

Immunocytochemistry - Anti-PUMA antibody (AB9643)

Immunocytochemical analysis of K562 cells labeling PUMA with ab9643 at 1 μg/mL. Cells were fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (AB9643)

Immunofluorescent analysis of 4% paraformaldehydefixed K562 cells labeling PUMA with ab9643 at 2 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing cytosol staining on K562 cells.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (AB9643)

ICC/IF image of ab9643 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9643, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• WB

AbReview3861****

Western blot - Anti-PUMA antibody (AB9643)

Blocked with 5% milk for 1 hour.

Incubated with the primary antibody for 18 hours.

All lanes:

Western blot - Anti-PUMA antibody (ab9643) at 1/1000 dilution

All lanes:

HeLa whole cell lysate

Secondary

All lanes:

Alexa Fluor 680-conjugated goat anti-rabbit IgG polyclonal at 1/1 dilution

Predicted band size: 21 kDa

Observed band size: 20 kDa,26 kDa,42 kDa,50 kDa

false

Exposure time: 5s

This image is courtesy of an anonymous Abreview

• WB

Supplier Data

Western blot - Anti-PUMA antibody (AB9643)

Beta-actin (1 μg/mL) and GAPDH (0.02 μg/mL). Incubation time : 1 hour at Room Temperature in 5% NFDM/TBST.

All lanes:

Western blot - Anti-PUMA antibody (ab9643) at 2 µg/mL

Lane 1:

HEK293 cells were transfected with control siRNAs at 15 µg

Lane 2:

HEK293 cells were transfected with PUMA siRNAs at 15 µg

Secondary

All lanes:

Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-PUMA antibody (AB9643)

All lanes:

Western blot - Anti-PUMA antibody (ab9643) at 2 µg/mL

Lane 1:

K562 cell lysate at 15 µg

Lane 2:

NIH3T3 cell lysate at 15 µg

Secondary

All lanes:

Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 21 kDa

false