Anti-NDUFB8 antibody [EPR15961]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling NDUFB8 antibody (red) with purified ab192878 at a dilution of 1/30. Goat anti rabbit IgG (Alexa Fluor^® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% tritonX-100 permeabilized HeLa cells labeling NDUFB8 with ab192878 at 1/50 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) (ab150077) secondary antibody at 1/400 dilution. Nuclear counter stained is DAPI (blue).

• IP

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Immunoprecipitation - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Western blot analysis of NDUFB8 immunoprecipitated from Human fetal heart lysate using ab192878 at 1/20 dilution. Lane 1 : Human fetal liver lysate. Lane 2 : PBS instead of Human fetal liver lysate.
Secondary : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-NDUFB8 antibody [EPR15961] (ab192878)

Predicted band size: 22 kDa

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• IHC-P

AbReview81843****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraformaldehyde-fixed mouse skeletal muscle tissue permeabilized with 0.3% Triton X100 in PBS stained with ab192878 at 1/50 dilution. Secondary antibody was Goat Anti-Rabbit IgG Antibody (H+L), Biotinylated at 1/3000 dilution. Samples were incubated with the primary antibody with 5% goat serum in 0.2% Triton X100 in PBS for 16 hours at 4°C. Blocking was done using 5% serum for 1 hour at 21°C. Heat mediated antigen retrieval with Tris/EDTA pH 9.0. ABC system used for signal amplification. Chromogenic reaction developed with DAB Substrate Kit (ab64238).Cell nuclei counterstained with Gill's hematoxylin I.

This image is courtesy of an Abreview submitted by Diego Perez Rodriguez

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

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

• WB

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Western blot - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/5000 dilution

Lane 1:

Human fetal liver lysate at 20 µg

Lane 2:

Human tonsil lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 22 kDa

Observed band size: 19 kDa

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• WB

Lab

Western blot - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Lanes 1 - 2 : Merged signal (red and green). Green - ab192878 observed at 19 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab192878 was shown to specifically react with NDUFB8 in wild-type HAP1 cells as signal was lost in NDUFB8 knockout cells. Wild-type and NDUFB8 knockout samples were subjected to SDS-PAGE. ab192878 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

NDUFB8 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 19 kDa

false

• WB

Supplier Data

Western blot - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/20000 dilution

All lanes:

Human fetal heart lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 22 kDa

Observed band size: 19 kDa

false

• WB

Supplier Data

Western blot - Anti-NDUFB8 antibody [EPR15961] (AB192878)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/5000 dilution

Lane 1:

C6 cell lysate at 10 µg

Lane 2:

RAW 264.7 cell lysate at 10 µg

Lane 3:

PC-12 cell lysate at 10 µg

Lane 4:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Predicted band size: 22 kDa

Observed band size: 19 kDa

false