Overview

  • Product name

    Anti-Cystathionase/CTH antibody
    See all Cystathionase/CTH primary antibodies
  • Description

    Rabbit polyclonal to Cystathionase/CTH
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse, Rabbit, Horse, Guinea pig, Cow, Cat, Dog, Pig, Drosophila melanogaster, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human Cystathionase/CTH aa 38-87 (N terminal).
    Sequence:

    VPPISLSTTFKQGAPGQHSGFEYSRSGNPTRNCLEKAVAALDGAKYCLAF


    (Peptide available as ab139491)

  • Positive control

    • Transfected 293T cell lysate.
  • General notes

     This product was previously labelled as Cystathionase

     

Properties

Applications

Our Abpromise guarantee covers the use of ab80643 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. Predicted molecular weight: 45 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function

    Catalyzes the last step in the transsulfuration pathway from methionine to cysteine. Has broad substrate specificity. Converts cystathionine to cysteine, ammonia and 2-oxobutanoate. Converts two cysteine molecules to lanthionine and hydrogen sulfide. Can also accept homocysteine as substrate. Specificity depends on the levels of the endogenous substrates. Generates the endogenous signaling molecule hydrogen sulfide (H2S), and so contributes to the regulation of blood pressure.
  • Pathway

    Amino-acid biosynthesis; L-cysteine biosynthesis; L-cysteine from L-homocysteine and L-serine: step 2/2.
  • Involvement in disease

    Defects in CTH are the cause of cystathioninuria (CSTNU) [MIM:219500]. It is an autosomal recessive phenotype characterized by abnormal accumulation of plasma cystathionine, leading to increased urinary excretion.
  • Sequence similarities

    Belongs to the trans-sulfuration enzymes family.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • CGL_HUMAN antibody
    • CTH antibody
    • cystathionase (cystathionine gamma-lyase) antibody
    • Cystathionine gamma lyase antibody
    • Cystathionine gamma-lyase antibody
    • Cysteine desulfhydrase antibody
    • Gamma cystathionase antibody
    • Gamma-cystathionase antibody
    • Homoserine deaminase antibody
    • Homoserine dehydratase antibody
    • MGC9471 antibody
    see all

Images

  • Anti-Cystathionase/CTH antibody (ab80643) at 1 µg/ml + transfected 293T cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa



    Gel concentration: 12%

References

This product has been referenced in:

  • Markó L  et al. Role of Cystathionine Gamma-Lyase in Immediate Renal Impairment and Inflammatory Response in Acute Ischemic Kidney Injury. Sci Rep 6:27517 (2016). WB . Read more (PubMed: 27273292) »
  • Rashid S  et al. Hydrogen sulphide-induced relaxation of porcine peripheral bronchioles. Br J Pharmacol 168:1902-10 (2013). Read more (PubMed: 23215842) »
See all 4 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.

The three cystathionase antibodies we have, aside from ab80643, have all been tested for western blotting but we do not have the corresponding blocking peptides. I recommend the monoclonal antibody ab54573, which is raised against full-length protein, not a peptide:

https://www.abcam.com/index.html?datasheet=54573

Please do not hesitate to contact us if you need any more advice or information

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Question

Hello, I made the following changes: - Loaded 30 ug protein per lane (lane labeled HUVEC 1 was prepared using RIPA lysis buffer and lysate was sufficiently concentrated to load 30 ug; lane labeled HUVEC 2 was prepared from a dilute lysate, using a TCA precipitation) - Used phosphatase inhibitors (Phosphatase Inhibitor Cocktail A or Phosphatase Inhibitor Cocktail B from Santa Cruz) in addition to the protease inhibitor cocktail during cell lysis - Used a 12% SDS-PAGE gel, as compared to the NuPAGE Bis-Tris precast gels used previously I obtained the following results: Blot 1: monoclonal CTH, with phosphatase inhibitors Used 1:250 ab54573. Observed two strong bands that may correspond to the appropriate MW of the two cystathionase variants (see attached). Blot 2: monoclonal CTH, without phosphatase inhibitors Used 1:250 ab54573. Appeared similar to Blot 1. Blot 3: polyclonal CTH, with phosphatase inhibitors Used 1:250 ab80643. Observed several strong bands that do not seem to correspond to the desired 39 or 45 kDa MW. Blot 4: polyclonal CTH with HeLa positive control, without phosphatase inhibitors Used 1:250 ab80643. Detected several strong bands, but slightly different compared to Blot 3. Blot 5: monoclonal CTH antibody from Novus/Abnova Used 1:500 Appeared similar to Blot 1 and 2. Thank you for all of the suggestions. I am very satisfied with the performance of the monoclonal antibody (ab54573), but still have not been able to troubleshoot the polyclonal antibody (a80643) successfully. I appreciate your continued assistance! Sincerely,

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Answer

Thank you very much for keeping me up to date with the performance of the antibodies. I am very happy to hear you could obtain satisfactory results from ab54573. On the other hand it is regrettable ab80643 didn’t perform accordingly. I appreciate the time you have spent in the laboratory and understand your concerns. I can suggest you have regrettably received a bad vial of ab80643. As the order for this antibody was placed within the last 6 months, it is still covered by our Abpromise guarantee, and if you wish to, I can send you a free replacement for this antibody (from a different lot) a reimbursement or a Credit Note in compensation. I am very sorry for the inconveniences, and I will be happy to assist you with this problem, and give a quick and efficient solution. Therefore I look forward to hearing from you letting me know how you would like to proceed. Have a nice day.

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Question

Hello, The antibodies that I have been having difficulty with are for detecting cystathionase in HUVECs: ab54573:, XX-XX-XXXX, lot #: XXXX ab80643:, XX-XX-XXXX, lot #: XXXX The pancreatic cells had been reported to have only low amounts of cystathionase, so we instead moved on to HUVECs which are known to produce cystathionase in detectable quantities. Staining the membrane and the gel showed good transfer of proteins to the membrane. When imaging the blot with secondary antibody, I was able to see strong signal from ab54573 but only for a band present at the top of the gel .I tried digesting the protein lysate at both higher and lower temperatures and still observed the same results. I have not tried a concentration of this antibody higher than 1:500. I wasn't able to see anything on the membrane with the ab80643, trying both 1:1000 and 1:500 concentrations of antibody .Blocking was done in 5% BSA/wash buffer. Since both these antibodies should detect the same protein, I am having the most difficulty determining why the results are so different (one has strong signal albeit in the wrong position, and the other does not). Are these antibodies sensitive to the phosphorylation of the protein? I will look into trying a phosphatase inhibitor with the lysate preparation. I would like to determine whether this lysate preparation is or is not appropriate for the cystathionase protein. Would it be possible to obtain a positive control to determine whether we can detect the protein? Thank you!

Read More
Answer

Thanks for your reply. We don’t have any evidence of the expression of cystathionase in HUVECs. If you have data to support the presence of the protein in the cell line, but not in high quantities, it would be better loading bigger amounts of sample per lane. Attending to the antibodies datasheets, for ab54573, K-562 cell lysate was used to detect Cystathionase. We do have in our catalogue K562 cell lysates that could be used as positive controls. The link to the search result is: https://www.abcam.com/index.html?pageconfig=searchresults&search=K 562&pt=12&sk=targ&sv=K562&sn=K562&l=1&fViewMore=1 For ab80643 the suggested positive control is Transfected 293T cell lysate. Unfortunately, we have 293T cell lysate in out catalogue, but not the transfected one, that would be the one recommended to work with. HeLa lysate can also be used as a positive control. There are some HeLa lysates available in the catalogue, if you wish to have a look: https://www.abcam.com/index.html?pageconfig=searchresults&search=HeLa&pt=12&sk=app&sv=69&sn=WB&l=2&fViewMore=1 I would encourage you to perform the WB again following the recommendations given (higher protein amount, using phosphatase inhibitors, concentrating the ab dilution as well as increasing the secondary concentration) and maybe using any of these lysates to check whether the problem comes from the antibody or from the protein expression level. In case you can’t notice any improvement, please contact me again and I’ll try to give a solution as soon as possible.

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Answer

Thank you for contacting us. Unless specified on the datasheet, all the Abcam antibodies are characterized under reducing conditions. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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