Complex III Subunit 5 Antibody (YA1865)

Complex III Subunit 5 Antibody (YA1865) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Complex III Subunit 5.

Complex III Subunit 5 is a component of the ubiquinol-cytochrome c oxidoreductase (also known as cytochrome b-c1 complex or complex III, CIII), a multisubunit transmembrane complex that is part of the mitochondrial electron transport chain, driving oxidative phosphorylation. The respiratory chain comprises three multisubunit complexes: succinate dehydrogenase (complex II, CII), ubiquinol-cytochrome c oxidoreductase (complex III, CIII), and cytochrome c oxidase (complex IV, CIV), which cooperate to transfer electrons derived from NADH and succinate to molecular oxygen, generating an electrochemical gradient across the inner membrane that powers transmembrane transport and ATP synthase. The cytochrome b-c1 complex catalyzes electron transfer from ubiquinol to cytochrome c, coupling this redox reaction to proton translocation across the mitochondrial inner membrane, with protons carried as hydrogens on the quinol. In the Q cycle, two protons are consumed from the matrix, four protons are released into the intermembrane space, and two electrons are transferred to cytochrome c. The Rieske protein, a catalytic core subunit containing a [2Fe-2S] iron-sulfur cluster, cycles between two conformational states during catalysis to transfer electrons from the quinol bound at the Q(0) site in cytochrome b to cytochrome c1 (by similar). Incorporation of UQCRFS1 is the penultimate step in complex III assembly. UQCRFS1 undergoes proteolytic processing after integration into the complex III dimer, with one fragment, subunit 9, corresponding to its mitochondrial targeting sequence (MTS). This processing is essential for proper insertion of UQCRFS1 into the complex III dimer, but persistent UQCRFS1-derived fragments may hinder the processing and assembly of newly imported UQCRFS1 into complex III, adversely affecting its structure and function.

P47985

Predicted band size: 30 kDa; Observed band size: 24 kDa

Affinity Purified

Non-conjugated

Unmodified

AB_3104235

Microbiology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:1000; IHC: 1:50-1:100; IF: 1:50-1:200; IP: 1:20

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  Western blot analysis of extracts from NIH/3T3 (lane 2(20μg) , K562 (lane 3(20μg) , C6 (lane 4(20μg),using Complex III Subunit 5 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
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  Immunocytochemistry analysis of Hela cells labeling Complex III Subunit 5 with Complex III Subunit 5 Antibody (HY-P82120) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Complex III Subunit 5 Antibody (HY-P82120)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
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  Immunocytochemistry analysis of HepG2 cells labeling Complex III Subunit 5 with Complex III Subunit 5 Antibody (HY-P82120) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Complex III Subunit 5 Antibody (HY-P82120) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded mouse liver tissue using Complex III Subunit 5 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded mouse liver tissue using Complex III Subunit 5 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-F, IHC-P, ICC/IF, IP

Liquid

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40% Glycerol, 0.01% Sodium azide and 0.05% BSA

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

A synthetic peptide of human RISP