SDHAF4 Antibody - BSA Free

Immunohistochemistry-Paraffin: SDHAF4 Antibody [NBP1-86324]

Immunohistochemistry-Paraffin: SDHAF4 Antibody [NBP1-86324] - Staining of human small intestine shows moderate to strong cytoplasmic positivity in glandular cells.

Immunohistochemistry-Paraffin: SDHAF4 Antibody [NBP1-86324]

Immunohistochemistry-Paraffin: SDHAF4 Antibody [NBP1-86324] - Staining of human fallopian tube shows moderate to strong cytoplasmic positivity in glandular cells.

Western Blot: SDHAF4 Antibody - BSA Free [NBP1-86324]

Analysis in control (vector only transfected HEK293T lysate) and C6orf57 over-expression lysate (Co-expressed with a C-terminal myc-DDK tag (~3.1 kDa) in mammalian HEK293T cells).

Western Blot: SDHAF4 Antibody - BSA Free [NBP1-86324] -

Structure of the SDHA-AF2-AF4 complex.a Ribbon diagram of the human SDHA-AF2-AF4 complex. SDHA is shown in gray, SDHAF2 is shown in cyan, and SDHAF4 is shown in orange. The covalent FAD is shown as a stick representation with carbons yellow, oxygens red, nitrogens blue, and phosphorous orange. The isoalloxazine functional group of the FAD is positioned between the flavin-binding domain of SDHA and the C-terminus of SDHAF4. Key interactions are shown in the inset. b Orientation of SDHAF2 and SDHAF4 in the complex. SDHA is shown as ribbons and SDHAF2 and SDHAF4 are shown as space-filling. The view is rotated 70 around the x-axis as compared to the view in (a). c Interactions between the C-terminus of SDHAF4 and the SDHA active site. The position of the conserved C-terminus is stabilized by interactions between SDHAF4D107 and SDHAF4F108 and SDHA active site residues SDHAR451 and SDHAH407. d Validation of SDHAF4 binding residues using mutagenesis. SDHAF4 containing the indicated C-terminal mutations was evaluated for the ability to displace SDHAF2 from the SDHA-AF2 complex. The assembly factors that remained bound to SDHA were visualized after the separation of the reaction on an SDS-PAGE gel. Mutations involved SDHAF4D107 (SDHAF4D107A, SDHAF4D107T and SDHAF4D107N), and SDHAF4F108 (SDHAF4F108A and SDHAF4 delta F108). The SDS-PAGE gel is representative of n = 4 independent experiments. ImageJ quantitation of SDHAF2 (teal) and SDHAF4 (brown) was used to calculate the percentage of each assembly bound to SDHA, as compared to a control (100%). This is expressed on the y-axis of each bar graph as mean values +/- SD. Bar graphs show mean values +/- SD, and statistics were calculated by paired two-tailed Student’s t-test. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-023-44563-7), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SDHAF4 Antibody - BSA Free [NBP1-86324] -

Pairwise interactions between SDHA, SDHAF2, and SDHAF4.The interaction of His-SDHA (2.6 uM) with SDHAF2 (6.4 uM) and SDHAF4 (8 uM) was evaluated by using a Ni-NTA pull-down assay. SDHA was tested in several of its forms: apo-SDHA, SDHA with bound non-covalent FAD, and holo-SDHA with covalently attached FAD. His6-SDHA was incubated with purified SDHAF2 and SDHAF4 as indicated and associated proteins were evaluated by SDS-PAGE. FAD was added at 75 uM and fumarate was added at 5 mM. Yellowish FAD fluorescence is observed when SDHA is covalently attached to FAD. ImageJ densitometry, shown at the bottom, was measured as arbitrary units (arb. units.) and used to evaluate the relative binding of SDHAF2 (teal) and SDHAF4 (brown) to SDHA. The y-axis on the densitometry quantitation expresses these as a ratio. a Input protein and pairwise interaction between SDHA and SDHAF2. (left) input SDHA, (right) interaction between SDHA and SDHAF2 in the presence of FAD and fumarate. Note that only after the addition of fumarate does the covalent bond between FAD and SDHA form (lane 3). b Pairwise interaction of SDHA and SDHAF4. c Interaction of SDHA with the assembly factors after incubation with both SDHAF2 and SDHAF4. d Displacement of SDHAF2 after purified holo-SDHA/SDHAF2 complex (2 uM) was incubated with SDHAF4. All Coomassie gels are representative of n = 4 independent experiments, bar graphs show mean values +/- SD, and statistics were done by paired two-tailed Student’s t-test. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-023-44563-7), licensed under a CC-BY license. Not internally tested by Novus Biologicals.