Anti-NDUFS3 antibody [EPR12781] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12781] - BSA and Azide free (AB240276)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling NDUFS3 with ab183733 at 1/50 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183733).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-NDUFS3 antibody [EPR12781] - BSA and Azide free (AB240276)

This data was developed using the same antibody clone in a different buffer formulation (ab183733).

Lanes 1- 4 : Merged signal (red and green). Green - ab183733 observed at 30 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab183733 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type HEK-293T and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183733 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFS3 antibody [EPR12781] (<a href='/ko/products/primary-antibodies/ndufs3-antibody-epr12781-ab183733'>ab183733</a>) at 1/10000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HL60 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• WB

Lab

Western blot - Anti-NDUFS3 antibody [EPR12781] - BSA and Azide free (AB240276)

This data was developed using the same antibody clone in a different buffer formulation (ab183733).

Lanes 1 - 2 : Merged signal (red and green). Green - ab183733 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab183733 was shown to react with NDUFS3 in HEK-293T wild-type cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab183733 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFS3 antibody [EPR12781] (<a href='/ko/products/primary-antibodies/ndufs3-antibody-epr12781-ab183733'>ab183733</a>) at 1/10000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

Predicted band size: 30 kDa

Observed band size: 27 kDa

false