Anti-Ndufs1 antibody [EPR11521(B)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling Ndufs1 with ab169540 at 1/50 dilution (2.6 ug/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic staining in Raji cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling Ndufs1 with ab169540 at 1/100 (1.29 ug/ml) followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney. The section was incubated with ab169540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 20 min.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling Ndufs1 with ab169540 at 1/20 dilution (1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

This data was developed using ab169540, the same antibody clone in a different buffer formulation. Ndufs1 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab169540 at 1/20 dilution (0.6ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab169540 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate Lane 2 : ab169540 IP in Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab169540 in Raji whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 7 seconds

All lanes:

Immunoprecipitation - Anti-Ndufs1 antibody [EPR11521(B)] (ab169540) at 1/30 dilution

All lanes:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 79 kDa

Observed band size: 75 kDa

false

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labelling Ndufs1 with ab169540 at 1/100 (1.29 ug/ml) followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney. The section was incubated with ab169540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 20 min.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labelling Ndufs1 with ab169540 at 1/100 (1.29 ug/ml) followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney. The section was incubated with ab169540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 20 min.

• WB

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Western blot - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

All lanes:

Western blot - Anti-Ndufs1 antibody [EPR11521(B)] (ab169540) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysat at 20 µg

Lane 3:

Mouse brain lysate at 20 µg

Lane 4:

Rat brain 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: 79 kDa

Observed band size: 75 kDa

false

Exposure time: 7s

• WB

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Western blot - Anti-Ndufs1 antibody [EPR11521(B)] (AB169540)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Ndufs1 antibody [EPR11521(B)] (ab169540) at 1/10000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) without mitochondria fraction cell lysate at 20 µg

Lane 2:

HepG2 mitochondria fraction cell 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: 79 kDa

Observed band size: 75 kDa

false