HRP Anti-SDHB antibody [21A11AE7]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-SDHB antibody [21A11AE7] (AB197903)

IHC image of SDHB staining in a section of formalin-fixed paraffin-embedded human normal colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab197903 at 1/100 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Lab

Western blot - HRP Anti-SDHB antibody [21A11AE7] (AB197903)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab197903 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-SDHB antibody [21A11AE7] (ab197903) at 1/5000 dilution

All lanes:

Human heart tissue lysate - mitochondrial extract (<a href='/en-us/products/unavailable/human-heart-tissue-lysate-mitochondrial-extract-ab110337'>ab110337</a>) at 5 µg

Predicted band size: 32 kDa

Observed band size: 32 kDa

true

Exposure time: 6s

• WB

Supplier Data

Western blot - HRP Anti-SDHB antibody [21A11AE7] (AB197903)

Lanes 1 - 3 : Merged signal (red and green). Green - ab197903 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab197903 was shown to specifically react with SDHB in wild-type HEK-293 cells as signal was lost in SDHB knockout cell line ab260860 (knockout cell lysate ab261652). Wild-type and SDHB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab197903 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 - HRP Anti-SDHB antibody [21A11AE7] (ab197903) at 1/5000 dilution

Lane 1:

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

Lane 2:

SDHB knockout HEK-293 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 whole cell lysate at 20 µg

Predicted band size: 32 kDa

Observed band size: 32 kDa

false