Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human heart tissue labeling SDHB with ab178423 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SDHB with ab178423 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of permeabilized HepG2 cells labeling SDHB with ab178423 at 1/10 dilution (red) compared with a rabbit IgG negative control (green).

• IP

Supplier Data

Immunoprecipitation - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation.

Western blot analysis on immunoprecipitation pellet from (1) A431 cell lysate or (2) 1X PBS (negative control) immunoprecipitated using ab178423 at 1/10 dilution. Detection utilized HRP-conjugated anti-rabbit IgG which preferentially detects the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-SDHB antibody [EPR13042(B)] (<a href='/en-us/products/primary-antibodies/sdhb-antibody-epr13042b-ab178423'>ab178423</a>)

Predicted band size: 32 kDa,76 kDa

Observed band size: 100 kDa,85 kDa

false

• WB

Lab

Western blot - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation. Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : SDHB knockout HEK-293 whole cell lysate
Lane 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lanes 1 - 3 : Merged signal (red and green). Green - ab178423 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab178423 was shown to recognize SDHB in wild-type HEK-293 cells as signal was lost at the expected MW in SDHB knockout cell line ab260860 (knockout cell lysate ab261652). Additional cross-reactive bands were observed in the wild-type and knockout cell lysate. Wild-type and SDHB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab178423 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/5000 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 [EPR13042(B)] (<a href='/en-us/products/primary-antibodies/sdhb-antibody-epr13042b-ab178423'>ab178423</a>) at 1/5000 dilution

Lane 1:

Wild-type 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 lysate (<a href='/en-us/products/cell-lysates/human-sdhb-knockout-hek-293-cell-lysate-ab261652'>ab261652</a>) at 20 µg

Lane 3:

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

Secondary

Lanes 1 - 3:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 32 kDa,130 kDa

false

• WB

Supplier Data

Western blot - Anti-SDHB antibody [EPR13042(B)] - BSA and Azide free (AB250046)

This data was developed using ab178423, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SDHB antibody [EPR13042(B)] (<a href='/en-us/products/primary-antibodies/sdhb-antibody-epr13042b-ab178423'>ab178423</a>) at 1/1000 dilution

Lane 1:

Human fetal liver tissue lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 32 kDa

false