Anti-Hexokinase II antibody [3D3]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [3D3] (AB104836)

ab104836 at 1/200 dilution staining Hexokinase II in human esophagus cancer by Immunohistochemistry, Paraffin-embedded tissue with DAB staining.

• IHC-P

AbReview32319****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [3D3] (AB104836)

Immunohistochemical analysis of human skin tissue, staining Hexokinase II with ab104836.

Tissue was fixed with paraformaldehyde and blocked with 10% BSA for 2 hours at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/250 in 5% NGS + 10% BSA in PBS) for 16 hours at 4°C. An HRP-conjugated goat anti-mouse polyclonal IgG (1/250) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Ms Hadas Pahima

• Flow Cyt

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Flow Cytometry - Anti-Hexokinase II antibody [3D3] (AB104836)

ab104836 staining Hexokinase II in K562 cells by Immunofluorescence (green) and negative control (purple).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [3D3] (AB104836)

ab104836 at 1/200 dilution staining Hexokinase II in human lung cancer by Immunohistochemistry, Paraffin-embedded tissue with DAB staining.

• WB

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Western blot - Anti-Hexokinase II antibody [3D3] (AB104836)

All lanes:

Western blot - Anti-Hexokinase II antibody [3D3] (ab104836) at 1/500 dilution

Lane 1:

Jurkat cell lysate

Lane 2:

HeLa cell lysate

Lane 3:

HEK293 cell lysate

Predicted band size: 102 kDa

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• WB

Lab

Western blot - Anti-Hexokinase II antibody [3D3] (AB104836)

Lanes 1 - 4 : Merged signal (red and green). Green - ab104836 observed at 102 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

ab104836 was shown to react with Hexokinase II in wild-type HEK-293T cells in western blot. The band observed in HK2 CRISPR/Cas9 edited sample lane below 102 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and HK2 CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab104836 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hexokinase II antibody [3D3] (ab104836) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HK2 CRISPR/Cas9 edited HEK-293T cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Predicted band size: 102 kDa

Observed band size: 102 kDa

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