Anti-SDHB antibody [EPR10880]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human mesenchymoma tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (AB175225)

Intracellular flow cytometric analysis of permeabilized Jurkat cellslabeling SDHB with unpurified ab175225 at a dilution of 1/10 (red) compared toa rabbit IgG negative control (green).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (AB175225)

Intracellular Flow Cytometry analysis ofA431 cells labelling SDHB with purified ab175225 at a dilution of 1/200 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^®488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human carotid paraganglioma tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

• IP

Lab

Immunoprecipitation - Anti-SDHB antibody [EPR10880] (AB175225)

ab175225 (purified) at a dilution of 1/60 immunoprecipitating SDHB in Jurkat whole cell lysate.

Lane 1 (input) : Jurkat whole cell lysate (10μg)

Lane 2 (+) : ab175225 + Jurkat whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab175225 in Jurkat whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-SDHB antibody [EPR10880] (ab175225)

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (AB175225)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-SDHB antibody [EPR10880] (AB175225)

Lanes 1 - 3 : Merged signal (red and green). Green - ab175225 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab175225 was shown to recognize SDHB in wild-type HEK 293 cells as signal was lost at the expected MW in SDHB knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SDHB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab175225 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/50000 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-SDHB antibody [EPR10880] (ab175225) at 1/50000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

SDHB knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SDHB knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-sdhb-knockout-hek-293-cell-line-ab260860'>ab260860</a>)

Lane 3:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-SDHB antibody [EPR10880] (AB175225)

All lanes:

Western blot - Anti-SDHB antibody [EPR10880] (ab175225) at 1/50000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

A431 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Fetal heart tissue lysate at 10 µg

Predicted band size: 32 kDa

false

• WB

Lab

Western blot - Anti-SDHB antibody [EPR10880] (AB175225)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SDHB antibody [EPR10880] (ab175225) at 1/100000 dilution

Lane 1:

HepG2 whole cell lysate at 10 µg

Lane 2:

Jurkat whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

• WB

Lab

Western blot - Anti-SDHB antibody [EPR10880] (AB175225)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SDHB antibody [EPR10880] (ab175225) at 1/100000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Rat brain tissue lysate at 10 µg

Lane 3:

Mouse spleen tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 32 kDa

false