Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using the same antibody clone in a different buffer formulation (ab192022). ab192022 staining NDUFS2 in wild-type Hap1 cells (top panel) and NDUFS2 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab192022 at 0.4μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human brain tissue sections labeling NDUFS2 using ab192022 at a 1/500 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain. Negative control uses PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

Lane 1 : 293T cell lysate was immunoprecipitated using ab192022 at a 1/40 dilution. Secondary used was anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at a 1/1500 dilution.

Lane 2 : PBS instead of 293T lysates.

All lanes:

Immunoprecipitation - Anti-NDUFS2 antibody [EPR16266] (<a href='/en-us/products/primary-antibodies/ndufs2-antibody-epr16266-ab192022'>ab192022</a>)

Predicted band size: 53 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat stomach tissue sections labeling NDUFS2 using ab192022 at a 1/500 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain. Negative control uses PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-NDUFS2 antibody [EPR16266] (<a href='/en-us/products/primary-antibodies/ndufs2-antibody-epr16266-ab192022'>ab192022</a>) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 3:

293T (Human embryonic kidney epithelial cell) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa

false

• WB

Lab

Western blot - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 whole cell lysate (20 Âμg)

Lane 2 : NDUFS2 knockout HAP1 whole cell lysate (20 Âμg)

Lane 3 : HeLa whole cell lysate (20 Âμg)

Lane 4 : Jurkat whole cell lysate (20 Âμg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab192022 observed at 48 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab192022 was shown to specifically react with NDUFS2 in wild-type HAP1 cells whilst signal was lost in NDUFS2 knockout cells. Wild-type and NDUFS2 knockout samples were subjected to SDS-PAGE. ab192022 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20,000 dilution respectively. Blots were developed 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 1/20,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFS2 antibody [EPR16266] (<a href='/en-us/products/primary-antibodies/ndufs2-antibody-epr16266-ab192022'>ab192022</a>)

Predicted band size: 53 kDa

false

• WB

Supplier Data

Western blot - Anti-NDUFS2 antibody [EPR16266] - BSA and Azide free (AB251104)

This data was developed using ab192022, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-NDUFS2 antibody [EPR16266] (<a href='/en-us/products/primary-antibodies/ndufs2-antibody-epr16266-ab192022'>ab192022</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat kidney lysate at 10 µg

Lane 6:

C6 cell lysate at 10 µg

Lane 7:

RAW264.7 cell lysate at 10 µg

Lane 8:

PC-12 cell lysate at 10 µg

Lane 9:

NIH3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa

Observed band size: 49 kDa

false