Overview

  • Product name

    Anti-Cytochrome b245 Light Chain/p22-phox antibody
    See all Cytochrome b245 Light Chain/p22-phox primary antibodies
  • Description

    Rabbit polyclonal to Cytochrome b245 Light Chain/p22-phox
  • Host species

    Rabbit
  • Specificity

    Please note, previous lots of this product were guaranteed in ICC/IF. We are having trouble reproducing new lots suitable for ICC/IF. Therefore this application has been removed and it no longer guaranteed.
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide corresponding to Cytochrome b245 Light Chain/p22-phox aa 100 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (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.

Target

  • Relevance

    Cytochrome 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 localization

    Cell 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

Images

  • All lanes : Anti-Cytochrome b245 Light Chain/p22-phox 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
    All lanes : 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.



    We hypothesize that the 21 kDa band represents the pro-form of Cytochrome b245 Light Chain/p22-phox, and the 19 kDa band represents the mature form.

  • Immunohistochemical analyses of p22phox using ab75941 (1/200) in normal adrenocortical tissue (b-c) and adrenal aldosterone-producing adenoma, (e-f) p22phox presents a gradient of expression in the entire adrenal cortex (100x), and its localization was restricted to the cell membrane (400x). Sections were incubated with primary antibody at 4°C overnight and with a HRP-conjugated secondary antibody for 30 minutes at 37°C.

  • IHC image of Cytochrome b245 Light Chain/p22-phox staining in a section of formalin-fixed paraffin-embedded normal human lung* 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 20mins. The section was then incubated with ab75941, 1ug/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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

References

This product has been referenced in:

  • Feillet-Coudray C  et al. Long-Term Measures of Dyslipidemia, Inflammation, and Oxidative Stress in Rats Fed a High-Fat/High-Fructose Diet. Lipids 54:81-97 (2019). Read more (PubMed: 30767221) »
  • Sandoval R  et al. TNF-a Increases Production of Reactive Oxygen Species through Cdk5 Activation in Nociceptive Neurons. Front Physiol 9:65 (2018). Read more (PubMed: 29467671) »
See all 9 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 14 2017

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

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

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% gel)
Loading amount
50 µg
Specification
skeletal muscle
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Single isolated FDB skeletal muscle fibres)
Permeabilization
Yes - 0.1% Triton X-100 (for 10 hours)
Specification
Single isolated FDB skeletal muscle fibres
Blocking step
BSA as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Dr. G.K Sakellariou

Verified customer

Submitted Sep 25 2012

Answer

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