Anti-CTHRC1 antibody (ab85739)

Reviews (2)Q&A (4)References (8)

Overview

  • Product name

    Anti-CTHRC1 antibody
    See all CTHRC1 primary antibodies

  • Description

    Rabbit polyclonal to CTHRC1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Fr, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow

  • Immunogen

    Synthetic peptide corresponding to Human CTHRC1 aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab101727)

  • Positive control

    • This antibody gave a positive signal in the following lysates: Brain (Human) Tissue Lysate; Heart (Human) Tissue Lysate; Blood Vessel: artery (Human) Membrane Lysate; HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate; HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab85739 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 20 kDa (predicted molecular weight: 26 kDa).
IHC-Fr Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • Western blot - Anti-CTHRC1 antibody (ab85739)
    Western blot - Anti-CTHRC1 antibody (ab85739)
    All lanes : Anti-CTHRC1 antibody (ab85739) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human heart tissue lysate - total protein (ab29431)
    Lane 3 : Human blood vessel: artery normal tissue lysate - membrane extract (ab28989)
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 26 kDa
    Observed band size: 20 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 minutes


    We predict that the 20 kDa band represents the cleaved form of CTHRC1. Abcam welcomes customer feedback.
  • Immunohistochemistry (Frozen sections) - Anti-CTHRC1 antibody (ab85739)
    Immunohistochemistry (Frozen sections) - Anti-CTHRC1 antibody (ab85739)This image is taken from an anonymous abreview.
    Ab85739 staining CTHRC1 on E8.5 mouse embryo sections by immunohistochemistry. The tissue was paraformaldehyde fixed and permeabilized in PBST prior to blocking in NOVOCASTRA protein block for 30 minutes at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 48 hour at 4°C. An Alexa Fluor 555 conjugated donkey anti-rabbit antibody, diluted 1/1000, was used as the secondary. Nuclear staining shown with DAPI.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-CTHRC1 antibody (ab85739)
    Immunocytochemistry/ Immunofluorescence - Anti-CTHRC1 antibody (ab85739)
    ICC/IF image of ab85739 stained HepG2 cells. The cells were 4% PFA 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 (ab85739, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • Western blot - Anti-CTHRC1 antibody (ab85739)
    Western blot - Anti-CTHRC1 antibody (ab85739)
    All lanes : Anti-CTHRC1 antibody (ab85739) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human heart tissue lysate - total protein (ab29431)
    Lane 3 : Human blood vessel: artery normal tissue lysate - membrane extract (ab28989)
    Lane 4 : Human brain tissue lysate - total protein (ab29466) with Human CTHRC1 peptide (ab101727) at 1 µg/ml
    Lane 5 : Human heart tissue lysate - total protein (ab29431) with Human CTHRC1 peptide (ab101727) at 1 µg/ml
    Lane 6 : Human blood vessel: artery normal tissue lysate - membrane extract (ab28989) with Human CTHRC1 peptide (ab101727) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 26 kDa
    Observed band size: 20 kDa why is the actual band size different from the predicted?


    Exposure time: 20 minutes
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTHRC1 antibody (ab85739)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTHRC1 antibody (ab85739)
    IHC image of CTHRC1 staining in human skin melanoma 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 ab85739, 5µg/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

This product has been referenced in:

  • Zheng M  et al. CTHRC1 overexpression promotes cervical carcinoma progression by activating the Wnt/PCP signaling pathway. Oncol Rep 41:1531-1538 (2019). Read more (PubMed: 30628714) »
  • Ni S  et al. CTHRC1 overexpression predicts poor survival and enhances epithelial-mesenchymal transition in colorectal cancer. Cancer Med 7:5643-5654 (2018). Read more (PubMed: 30302922) »
See all 8 Publications for this product

Publishing research using ab85739? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-4 of 4 Q&A

Question

We are interested in purchasing your anti-CTHRC1 antibody (Cat. No. ab85739) but we have a few questions we would like to have addressed:

1) Is the antibody provided free of carrier protein, -SH, -NH2 and ready for conjugation with HRP?
2) We are interested in using the antibody for WB and IHC-P applications. Will the working antibody dilutions for these applications be reasonably sufficient (at least 1/500) for many blots and slides?
3) Is it possible for us to request that the antibody be HRP-conjugated at Abcam prior to shipping? We would be very interested in this service.

Thank you in advance for your assistance.

Read More
Answer

Thank you for contacting Abcam regarding ab85739.


This antibody is currently provided in a buffer containing BSA and Sodium Azide, but at concentrations that would not inhibit a typical conjugation reaction. Therefore, it is suitable for conjugation as formulated.


Regarding working dilutions, for WB we recommend 1:500 and for IHC we recommend 1:100. These are suggested starting dilutions and would require further optimization with your samples.


If you are still interested in purchasing this product, I would be happy to provide additional information regarding our conjugation services.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

Read More

Question

I brought a product (rabbit polyclonal to CTHRC1, ab85739) from your company.
In the figure and instructions, you mentioned that the antibody detected a band of approximatelt 20 kDa.
However, the molecular mass of the protein is around 26 kDa.
My question is,have you performed some experiments to prove that the band detected was indeed CTHRC1?

Read More
Answer

Thank you for contacting us and sorry for the delay in getting back to you.

I now have further information to share with you in regards to the anti-CTHRC1 antibody (ab85739). In order to try to validate the specificity of the antibody we have now performed Western blots including the blocking peptide ab101727 at a concentration of 1 ug/mL. The application of this peptide resulted in the loss of the detected band at ˜20 kDa. This result can be viewed on the datasheet of this antibody:

https://www.abcam.com/CTHRC1-antibody-ab85739.html

Additionally, we now also have further IHC testing data that show a very strong specific signals of the notochord of E8.5 mouse embryo sections as is consistent with the expected localisation of CTHRC1. This data can also be viewed from the datasheet, or from the following link:

https://www.abcam.com/index.html?datasheet=85739&tab=abreviews&intabreviewid=30160

I hope this information has been of help. If you have any further questions or concerns please do not hesitate to contact us again.

Read More

Question

I brought a product (rabbit polyclonal to CTHRC1, ab85739) from your company.
In the figure and instructions, you mentioned that the antibody detected a band of approximatelt 20 kDa.

However, the molecular mass of the protein is around 26 kDa.
My question is,have you performed some experiments to prove that the band detected was indeed CTHRC1?

Read More
Answer

Thank you for contacting us and making us aware of your concerns over the anti-CTHRC1 antibody (ab85739).

Unfortunately,wedo not have any furthertesting information than what is shown on the public datasheet for this antibody. We are currently setting up a Western blot usingthe immunisingpeptide asablocking peptide to validate the specific binding.

We thank you for bringing this to our attention and will let you know what the results ofthe further validation experiments are.

I am sorry that I could not be of more help in this instance. If you have any further questions please do not hesitate to contact us again.

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Question

I was just planning to ask for a replacement for this antibody,because we were not able to detect either recombinant human CTHRC1 protein or intracellular or secreted human CTHRC1 in Western blot with this antibody. Would it be possible to get the anti-cthrc1 antibody (ab85739) as a replacement? Best regards,

Read More
Answer

Thank you for contacting Abcam Technical Team and I am very sorry to hear that you are having problems with this product.   I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.   This is to let you know that I have placed a new order for you - for one vial of ab85739 as a free of charge replacement (exchange for the original ab72527) and the new order number is 986578.     I hope this alternative product will work as it is expected, and please do let me know how you are getting on with this product.  

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