Anti-Cytochrome b245 Light Chain antibody (ab75941)

Overview

  • Product nameAnti-Cytochrome b245 Light Chain antibody
    See all Cytochrome b245 Light Chain primary antibodies
  • Description
    Rabbit polyclonal to Cytochrome b245 Light Chain
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus Cytochrome b245 Light Chain.

    (Peptide available as ab86646.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HepG2 Whole Cell, Mouse Pancreas Tissue, Mouse Spleen Tissue. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

Properties

Applications

Our Abpromise guarantee covers the use of ab75941 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21 kDa).
IHC-P Use a concentration of 1 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • RelevanceCytochrome b245 Light Chain/p22phox is a critical component of the membrane-bound oxidase complex that generates superoxide. It complexes with NOX3 to form a functional NADPH oxidase.
  • Cellular localizationCell Membrane
  • Database links
  • Alternative names
    • CYBA antibody
    • Cytochrome b 245 alpha polypeptide antibody
    • Cytochrome b 245 light chain antibody
    • Cytochrome b alpha polypeptide antibody
    • Cytochrome b(558) alpha chain antibody
    • Cytochrome b(558) alpha subunit antibody
    • Cytochrome b558 subunit alpha antibody
    • Flavocytochrome b 558 alpha polypeptide antibody
    • Neutrophil cytochrome b 22 kDa polypeptide antibody
    • p22 phagocyte B-cytochrome antibody
    • p22-phox antibody
    • p22phox antibody
    • Superoxide-generating NADPH oxidase light chain subunit antibody
    see all

Anti-Cytochrome b245 Light Chain antibody images

  • All lanes : Anti-Cytochrome b245 Light Chain antibody (ab75941) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : Pancreas (Mouse) Tissue Lysate
    Lane 3 : Spleen (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 21 kDa
    Additional bands at : 19 kDa (possible cleavage fragment),40 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab75941 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75941, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa cells at 5µg/ml.
  • IHC image of Cytochrome b245 Light Chain staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75941, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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.

References for Anti-Cytochrome b245 Light Chain antibody (ab75941)

This product has been referenced in:
  • Murray TV  et al. NADPH oxidase 4 regulates cardiomyocyte differentiation via redox activation of c-Jun protein and the cis-regulation of GATA-4 gene transcription. J Biol Chem 288:15745-59 (2013). WB ; Mouse . Read more (PubMed: 23589292) »
  • Sakellariou G  et al. Studies of mitochondrial and non-mitochondrial sources implicate NADPH oxidase(s) in the increased skeletal muscle superoxide generation that occurs during contractile activity. Antioxid Redox Signal : (2012). WB, ICC/IF ; Mouse . Read more (PubMed: 23050834) »

See all 3 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (15%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 02 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (15%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 02 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (skeletal muscle)
Loading amount 50 µg
Specification skeletal muscle
Gel Running Conditions Reduced Denaturing (15% gel)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Single isolated FDB skeletal muscle fibres)
Specification Single isolated FDB skeletal muscle fibres
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100 (for 10 hours)
Blocking step BSA as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Dr. G.K Sakellariou

Verified customer

Submitted Sep 25 2012

Vielen Dank für Ihre Anfrage. Gegen die genannten NADPH-Oxidase-Komponenten haben wir folgende Antikörper bzw. Proteine in unserem Katalog: Click here (or use the following: http://www.abcam.com/index.html?datasheet=109366). Click here (or use the...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"