Anti-MTHFD2 antibody [4G7-2G3]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD2 antibody [4G7-2G3] (AB56772)

MTHFD2 antibody (ab56772) used in immunohistochemistry at 3μg/ml on formalin fixed and paraffin embedded human lymphoma tissue.

• Flow Cyt

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Flow Cytometry - Anti-MTHFD2 antibody [4G7-2G3] (AB56772)

Overlay histogram showing HepG2 cells stained with ab56772 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56772, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using the ascites version of the product.

• WB

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Western blot - Anti-MTHFD2 antibody [4G7-2G3] (AB56772)

MTHFD2 antibody (ab56772) at 1μg/lane + HepG2 cell lysate at 25μg/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)

Predicted band size: 38 kDa,42 kDa,43 kDa

Observed band size: 42 kDa

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• WB

Lab

Western blot - Anti-MTHFD2 antibody [4G7-2G3] (AB56772)

Lanes 1-4 : Merged signal (red and green). Green - ab56772 observed at 36 kDa. Red - loading control ab52901 observed at kDa.

ab56772 Anti-MTHFD2 antibody was shown to specifically react with MTHFD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266383 (knockout cell lysate ab258528) was used. Wild-type and MTHFD2 knockout samples were subjected to SDS-PAGE. ab56772 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MTHFD2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MTHFD2 knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-mthfd2-knockout-hek-293t-cell-line-ab266383'>ab266383</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 38 kDa

Observed band size: 36 kDa

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