Overview

  • Product name
    Anti-Cytochrome P450 Reductase antibody
    See all Cytochrome P450 Reductase primary antibodies
  • Description
    Rabbit polyclonal to Cytochrome P450 Reductase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Guinea pig, Hamster, Cow, Dog, Human, Pig, Monkey
  • Immunogen

    Purified rat liver NADPH-Cytochrome P450 reductase.

  • Positive control
    • Rat Liver NADPH Cytochrome P450 Reductase Protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab13513 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
ICC 1/500 - 1/1000.
IP Use a concentration of 5 µg/ml.
WB 1/1000. Detects a band of approximately 78 kDa.
IHC-P 1/500.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function
    This enzyme is required for electron transfer from NADP to cytochrome P450 in microsomes. It can also provide electron transfer to heme oxygenase and cytochrome B5.
  • Involvement in disease
    Defects in POR are the cause of adrenal hyperplasia variant type (AHV) [MIM:201750]; also known as Antley-Bixler syndrome-like phenotype with disordered steroidogenesis. AHV is a rare variant of congenital adrenal hyperplasia. It is an autosomal recessive disorder with apparent combined P450C17 and P450C21 deficiency. Affected girls are born with ambiguous genitalia, but their circulating androgens are low and virilization does not progress. Conversely, affected boys are sometimes born undermasculinized. Boys and girls can also present with bone malformations, in some cases resembling the pattern seen in patients with Antley-Bixler syndrome.
    Defects in POR are a cause of isolated disordered steroidogenesis (IDS) [MIM:201750].
  • Sequence similarities
    In the C-terminal section; belongs to the flavoprotein pyridine nucleotide cytochrome reductase family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Cellular localization
    Endoplasmic reticulum membrane. Anchored to the ER membrane by its N-terminal hydrophobic region.
  • Information by UniProt
  • Database links
  • Alternative names
    • CPR antibody
    • CYPOR antibody
    • Cytochrome p450 oxidoreductase antibody
    • DKFZp686G04235 antibody
    • FLJ26468 antibody
    • NADPH Cytochrome P450 Reductase antibody
    • NADPH dependent cytochrome P450 reductase antibody
    • NADPH--cytochrome P450 reductase antibody
    • NCPR_HUMAN antibody
    • P450 (cytochrome) oxidoreductase antibody
    • P450 Cytochrome Oxidoreductase antibody
    • P450R antibody
    • por antibody
    see all

Images

  • Anti-Cytochrome P450 Reductase antibody (ab13513) at 1/2000 dilution + Cell lysates prepared from rat PC12 cells


    Western blot analysis of rat PC12 cell lysate, probed with ab13513.
  • ICC/IF image of ab13513 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab13513, 10µg/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.

  • Cytochrome P450 Reductase was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Cytochrome P450 Reductase and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab13513.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 77kDa: Cytochrome P450 Reductase

References

This product has been referenced in:
  • Yao Y  et al. Effects and mechanism of amyloid ß1-42 on mitochondria in astrocytes. Mol Med Rep 17:6997-7004 (2018). Read more (PubMed: 29568933) »
  • Barzi M  et al. A novel humanized mouse lacking murine P450 oxidoreductase for studying human drug metabolism. Nat Commun 8:39 (2017). WB, IHC-Fr . Read more (PubMed: 28659616) »
See all 15 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (transfection of GFP tagged Human CYPred)
Loading amount
40000 cells
Specification
transfection of GFP tagged Human CYPred
Treatment
1x laemmli buffer, 10 min boiling,
Gel Running Conditions
Non-reduced Denaturing (4-20% TGX biorad)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Ms. Anette Orjuela

Verified customer

Submitted Feb 23 2012

Answer

Thank you for bringing this to our attention and for sending your protocol and IHC images. I am sorry to see that the antibody is giving you trouble. It is unlikely, but possible, that the wrong antibody was delivered to you. We have not received other complaints about the antibody so I do not think there was a labeling mix-up. The antibody may itself be defective, at least with these mouse samples. I am wondering if the glutaraldehyde that you include in the fixative might be creating some of the background but your negative control appears relatively clean. Some people will add glycine at 0.3M to the blocking solution to quench unreacted aldehydes. In any case, there is no staining of the ER, as far as I can tell. Can you tell me how long your samples were fixed, how much antibody you applied (concentration or dilution), and if you are confident that the secondary antibody is effective? For a replacement or refund or credit, I will also need the order reference number, either Abcam's or the PO number. I look forward to your reply and to resolving this issue.

Read More

Question
Answer

Thank you for contacting us with your question. Ab13513 is provided as whole antiserum and is not a purified antibody. Some of our other cytochrome P450 reductase antibodies are affinity purified (such as ab39995) and we also have purification kits that can be used to isolate the IgG molecules from the antiserum- https://www.abcam.com/Antibody-Serum-Purification-kit-1-purification-ab109208.html I hope this information will be useful in determining whether this antibody is appropriate for your studies. If you have any further questions or need any other information please let me know.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver whole homogenate)
Loading amount
10 µg
Specification
Liver whole homogenate
Gel Running Conditions
Reduced Denaturing (12 %)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 21 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (BS2B Human Lung Cells)
Loading amount
30 µg
Specification
BS2B Human Lung Cells
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5%

Mr. Adam Szadkowski

Verified customer

Submitted Jun 15 2007

Question
Answer

Thank you for contacting us. For ab23677, LTA4 hydrolase, I suggest either a neutrophil or monocyte lysate. We carry THP1 lysate, ab7913, which is a human monocytic leukemia. For a negative control, LTA4 was detected in epidermal keratinocytes and endothelial cells but not whole skin in the following paper: Leukotriene A4 hydrolase in human skin. J Invest Dermatol. 1994 Feb;102(2):253-7. PMID: 8106755 Our lysate ab30166 may be suitable as a negative control but I cannot guarantee you will not see some reactivity. For the cytochrome P450 enzymes, I suggest liver lysates. A positive control for ab4236 is human liver: ab29889. For ab13513 and ab3573, rat liver lysate ab29413 will be appropriate. I am reluctant to recommend a negative control since information regarding absence of expression is difficult to come by but since CP450 is associated with the ER, cytoplasmic fractions of liver lysates (which do not contain membrane) may be useful. For human liver, this would be ab29875, and for rat liver ab29409. You should also have a look at the references listed on the datasheets to see how other researchers have used the antibodies, though the references for the LTA4 are general and not specific to the antibody. I hope this helps. Please do not hesitate to contact us if you have any other questions.

Read More

Question
Answer

Thank you for your enquiry. I was able to obtain a Western blot image, which I have added to our datasheet. The antibody was used in WB in this paper, http://cancerres.aacrjournals.org/cgi/content/full/60/13/3550 I hope this information helps. Please do not hesitate to contact us if you need anything further.

Read More

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

Sign up