Overview

  • Product name
  • Description
    Rabbit polyclonal to CPS1
  • Host species
    Rabbit
  • Specificity
    When the antibody is used at 1/2000 some lower molecular weight bands appear in COS-7 cells, the extent of which is significantly reduced when the antibody is used at 1/10,000.
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Full length native protein (purified) (Rat).

  • Positive control
    • Rat liver
  • General notes
    OMIM reference: 114010 Alternative name: carbamoylphosphate synthetase 1, CAD

Properties

Applications

Our Abpromise guarantee covers the use of ab3682 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
IHC-Fr 1/100.
IHC-P 1/500.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

Target

  • Function
    Involved in the urea cycle of ureotelic animals where the enzyme plays an important role in removing excess ammonia from the cell.
  • Tissue specificity
    Primarily in the liver and small intestine.
  • Involvement in disease
    Defects in CPS1 are the cause of carbamoyl phosphate synthetase 1 deficiency (CPS1D) [MIM:237300]. CPS1D is an autosomal recessive disorder of the urea cycle causing hyperammonemia. Clinical features include protein intolerance, intermittent ataxia, seizures, lethargy, developmental delay and mental retardation.
    Note=Genetic variations in CPS1 influence the availability of precursors for nitric oxide (NO) synthesis and play a role in clinical situations where endogenous NO production is critically important, such as neonatal pulmonary hypertension, increased pulmonary artery pressure following surgical repair of congenital heart defects or hepatovenocclusive disease following bone marrow transplantation. Infants with neonatal pulmonary hypertension homozygous for Thr-1406 have lower L-arginine concentrations than neonates homozygous for Asn-1406.
  • Sequence similarities
    Contains 2 ATP-grasp domains.
    Contains 1 glutamine amidotransferase type-1 domain.
  • Domain
    The type-1 glutamine amidotransferase domain is defective.
  • Cellular localization
    Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • Carbamoyl phosphate synthase [ammonia] antibody
    • Carbamoyl phosphate synthase [ammonia] mitochondrial antibody
    • Carbamoyl phosphate synthase antibody
    • Carbamoyl phosphate synthetase 1 antibody
    • Carbamoyl phosphate synthetase 1 mitochondrial antibody
    • Carbamoyl phosphate synthetase I antibody
    • Carbamoyl-phosphate synthase [ammonia] antibody
    • Carbamoyl-phosphate synthetase I antibody
    • Carbamoylphosphate synthase antibody
    • Carbamoylphosphate synthetase 1 antibody
    • Carbamoylphosphate synthetase I antibody
    • CPS 1 antibody
    • Cps1 antibody
    • CPSase 1 antibody
    • CPSase I antibody
    • CPSASE1 antibody
    • CPSM_HUMAN antibody
    • mitochondrial antibody
    • MS738 antibody
    see all

Images

  • Lane 1: 5 ul of 2ml lysate cos-7 cells transfected with full length CPS 1. Before IP
    Lane 2: cos-7 cells transfected with full length CPS 1. IP with 5 ul of pre-immune serum
    Lane 3: cos-7 cells transfected with full length CPS 1. IP with 2 ul of polyclonal antibody anti CPS 1 (ab3682)
    Lane 4: cos-7 cells transfected with full length CPS 1. IP with 5 ul of polyclonal antibody anti CPS 1 (ab3682)
    Lane 5: 5 ul of 2ml lysate cos-7 cells transfected with empty vector (pCMV5-tr). Before IP
    Lane 6: cos-7 cells transfected with empty vector (pCMV5-tr). IP with 5 ul of polyclonal antibody anti CPS 1 (ab3682)
    Lane 7: 5 ul of 2ml lysate untransfected cos-7 cells. Before IP
    Lane 8: untransfected cos-7 cells. IP with 5 ul of polyclonal antibody anti CPS 1 (ab3682) Ab3682 can IP CPS1 transiently expressed in COS-7 cells.

    Lane 1: 5 ul of 2ml lysate cos-7 cells transfected with full length CPS 1. Before IP
    Lane 2: cos-7 cells transfected with full length CPS 1. IP
  • CPS1 - Liver Mitochondrial Marker was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to CPS1 - Liver Mitochondrial Marker (ab3682) 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, Hela whole cell extract 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 ab3682.
    Secondary: Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 165kDa: CPS1.
  • ab3682 staining CPS1 in mouse hepatic tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent paraformaldehyde fixation before heat mediated antigen retrieval in Citrate buffer and then blocked in maleate blocking solution for 30 minutes at 22°C. The primary antibody was diluted 1/500 and incubated with sample in maleate buffer blocking solution for 16 hours at 22°C. A Biotin conjugated donkey polyclonal to rabbit IgG was used at dilution at 1/250 as secondary antibody.

    See Abreview

  • ab3682 staining CPS1 in mouse liver tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde and blocking with 100% maleate blocking solution at 220C for 30 minutes was performed. The sample was incubated with primary antibody (1/100) at 220C for 16 hours. An Alexa Fluor®488-conjugated donkey polyclonal to rabbit IgG was used undiluted as a secondary antibody. The image demonstrates CPS1 in green, nuclei stained blue with DAPI.

    See Abreview

  • CPS-1 localisation is consistent with the reticulated pattern of liver mitonchondria.

    Rabbit polyclonal to CPS-1 (ab3682), at 1/5000 on COS-7 cells transiently transfected with pCMV5-CPS 1 plasmid (Corvi, M.M and Berthiaume, L.G. unpublished data).

    Secondary: donkey anti-rabbit (Texas Red).

  • Western blot using Rabbit polyclonal to CPS 1 (ab3682). All lanes loaded with 100 ng of purified CPS 1
    Lane 1: ab3682 1/500
    Lane 2: ab3682 1/1000
    Lane 3: ab3682 1/2000 Western blot using Rabbit polyclonal to CPS 1 (ab3682).

    All lanes loaded with 100 ng of purified CPS 1

    Lane 1: ab3682 1/500
    Lane 2: ab3682 1/1000
    Lane 3: ab3682 1/2000

References

This product has been referenced in:
  • Kang YBA  et al. Metabolic Patterning on a Chip: Towards in vitro Liver Zonation of Primary Rat and Human Hepatocytes. Sci Rep 8:8951 (2018). ICC/IF . Read more (PubMed: 29895900) »
  • Wang T  et al. L-leucine stimulates glutamate dehydrogenase activity and glutamate synthesis by regulating mTORC1/SIRT4 pathway in pig liver. Anim Nutr 4:329-337 (2018). Read more (PubMed: 30175263) »
See all 16 Publications for this product

Customer reviews and Q&As

1-3 of 3 Q&A

Question
Answer

Vielen Dank für Ihren Anruf heute Morgen.
Ich habe mir die Beschreibung des IHC-P Bildes nochmals angeschaut und muss ihnen leider mitteilen, dass ich Ihnen keine genauen Angaben über dem Maleate Puffer machen kann, da dieses Bild und die Beschreibung nicht aus unsrem Labor stammt. Eine Kundin hat einen Abreview als Feedback mi t diesem Bild eingeschickt.
https://www.abcam.com/index.html?datasheet=3682&tab=abreviews&intabreviewid=14931
Ich habe allerding Rezepte für Melate-Puffer gefunden:
http://www.nrims.harvard.edu/protocols/Uranyl_Acetate_Stain_in_Maleate_Buffer.pdf
http://www.ou.edu/research/electron/bmz5364/maleate.html
Ich hoffe, diese helfen Ihnen weiter und wünsche Ihnen viel Erfolg mit Ihren Experimenten.

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Question
Answer

Thank you for your enquiry. This antibody has been tested in both Western blot and IP, but only with rat. Unfortunately, we can not guarantee that this product will work in mouse. If you decide to use it in those applications with mouse samples, please consider submitting an Abreview to let us know how it went and also to earn Abpoints. Abpoints can be used towards various promotions including Amazon.com gift certificates and credit towards future antibody purchases. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry. As far as I am aware, this antibody (ab3682) has not been tested in wood frog Rana sylvatica. We are therefore not able to guarantee this antibody will work in this species. However, if you have the sequences to both rat (to which it was raised in) and wood frog, then it may be worth doing a BLAST (sequence homology analysis) search to see how homologous the proteins are. This antibody is a polyclonal to the full length native protein from rat. As a polyclonal, the antibodies it contains will therefore recognise many epitopes on the whole length of the protein. If you decide to use this antibody, I would be very pleased to hear how you get on! We always encourage customers to give us feedback if they have used an antibody in a new species or new application,whether positive or negative, and make all product information available to researchers. If you send us an Abreview, we can offer you from 50 up to 300 Abcam points (Abpoints) based on submission time and type of application. The less characterised the product you review, the more Abpoints you will receive. Furthermore, you get 100 extra Abpoints for submitting an image. These Abpoints can then be claimed back as rewards. I hope this helps. Please get back to me if you have any more questions or need any more advice. Good luck with your interesting research!

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