Anti-PCK2 antibody [RP23040240]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PCK2 antibody [RP23040240] (AB308126)

For immunofluorescence analysis, HepG2 cells were fixed and permeabilized for detection of endogenous PCK2 using ab308126 at 5 µg/mL) and labeled with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution. Panel a) shows representative cells that were stained for detection and localization of PCK2 protein (green) Panel b) is stained for nuclei (blue) using DAPI Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin at 1/300 dilution Panel d) is a composite image of Panels a, b and c clearly demonstrating Mitochondrial localisation of PCK2 Panel e) represents control cells with no primary antibody to assess background The images were captured at 60X magnification

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PCK2 antibody [RP23040240] (AB308126)

For immunofluorescence analysis, HepG2 cells were fixed and permeabilized for detection of endogenous PCK2 using ab308126 at 5 µg/mL and labeled with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution. Panel a) shows representative cells that were stained for detection and localization of PCK2 protein (green) Panel b) is stained for nuclei (blue) using DAPI Panel c) represents mitochondrial staining using a red mitochondria stain (MitoTracker) Panel d) is a composite image of Panels a, b and c clearly demonstrating co-localization of PCK2 with MitoTracker which specifically stains mitochondria Panel e) represents control cells with no primary antibody to assess background The images were captured at 60X magnification

• WB

Supplier Data

Western blot - Anti-PCK2 antibody [RP23040240] (AB308126)

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), A549 (Lane 2), HeLa (Lane 3), HT-29 (Lane 4), MDA-MB-231 (Lane 5), HEK-293 (Lane 6), A-431 (Lane 7), Jurkat (Lane 8) and C2C12 (Lane 9). The blots were probed with ab308126 at 2.5 µg/mL and detected by chemiluminescence using a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution. A 70 kDa band corresponding to PCK2 was observed across the 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 (ECL).

All lanes:

Western blot - Anti-PCK2 antibody [RP23040240] (ab308126) at 2.5 µg/mL

Lane 1:

HepG2 cell lysate at 30 µg

Lane 2:

A549 cell lysate at 30 µg

Lane 3:

HeLa cell lysate at 30 µg

Lane 4:

HT-29 cell lysate at 30 µg

Lane 5:

MDA-MB-231 cell lysate at 30 µg

Lane 6:

HEK-293 cell lysate at 30 µg

Lane 7:

A431 cell lysate at 30 µg

Lane 8:

Jurkat cell lysate at 30 µg

Lane 9:

C2C12 cell lysate at 30 µg

Secondary

All lanes:

HRP conjugated Goat anti-Rabbit IgG (H+L) Secondary Antibody at 1/4000 dilution

Observed band size: 70 kDa

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