Anti-SDHA antibody [EPR9043(B)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

Immunohistochemichal analysis of paraffin embedded human testis tissue labelling SDHA with ab137040 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

Immunohistochemichal analysis of paraffin embedded human kidney tissue labelling SDHA with ab137040 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 staining SDHA in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 staining SDHAin the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/500 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 staining SDHA in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab137040 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1 : Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2 : Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (AB137040)

Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling SDHA with ab137040 at 1/100 dilution.

• IP

Lab

Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 immunoprecipitating SDHA. 10μg of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1 : HeLa whole cell lysate (10ug)
Lane 2 : ab137040 IP in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab137040 in HeLa whole cell lysate

All lanes:

Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Predicted band size: 72 kDa

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• IP

Lab

Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 immunoprecipitating SDHA. 10μg of Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1 : Jurkat whole cell lysate 10ug
Lane 2 : ab137040 IP in Jurkat whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab137040 in Jurkat whole cell lysate

All lanes:

Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Predicted band size: 72 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 staining SDHA in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (AB137040)

ab137040 staining SDHA in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• WB

Supplier Data

Western blot - Anti-SDHA antibody [EPR9043(B)] (AB137040)

Lanes 1 - 4 : Merged signal (red and green). Green - ab137040 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab137040 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cell line ab261853 (knockout cell lysate ab261657). Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab137040 and ab8245 (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-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 2:

Western blot - Human SDHA knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-sdha-knockout-hek-293-cell-line-ab261853'>ab261853</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

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

Predicted band size: 72 kDa

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• WB

Lab

Western blot - Anti-SDHA antibody [EPR9043(B)] (AB137040)

Blocking buffer : 5% NFDM /TBST

Diluting buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/5000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma) whole cell lysate at 20 µg

Lane 3:

Jurkat (human acute T cell leukemia) whole cell lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Rat brain tissue lysate at 20 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: 72 kDa

Observed band size: 72 kDa

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• WB

Unknown

Western blot - Anti-SDHA antibody [EPR9043(B)] (AB137040)

All lanes:

Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

HT1080 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat anti Rabbit IgG at 1/2000 dilution

Predicted band size: 72 kDa

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