Overview

  • Product name

    Anti-Liver Carboxylesterase 1/CES1 antibody
    See all Liver Carboxylesterase 1/CES1 primary antibodies
  • Description

    Rabbit polyclonal to Liver Carboxylesterase 1/CES1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse, Baboon, Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Human Liver Carboxylesterase 1/CES1 aa 500 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab66527, ab66528)

  • Positive control

    • Recombinant Human Liver Carboxylesterase 1/CES1 protein (ab114347) can be used as a positive control in WB. This antibody gave a positive signal in the following Lysates: HepG2 Whole Cell THP1 Whole Cell Mouse Liver Tissue Rat Liver Tissue This antibody gave a positive result in IHC in the following FFPE tissue: Human normal liver.
  • General notes

     This product was previously labelled as Liver Carboxylesterase 1

     

Properties

Applications

Our Abpromise guarantee covers the use of ab45957 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-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function

    Involved in the detoxification of xenobiotics and in the activation of ester and amide prodrugs. Hydrolyzes aromatic and aliphatic esters, but has no catalytic activity toward amides or a fatty acyl-CoA ester. Hydrolyzes the methyl ester group of cocaine to form benzoylecgonine. Catalyzes the transesterification of cocaine to form cocaethylene. Displays fatty acid ethyl ester synthase activity, catalyzing the ethyl esterification of oleic acid to ethyloleate.
  • Tissue specificity

    Expressed predominantly in liver with lower levels in heart and lung.
  • Sequence similarities

    Belongs to the type-B carboxylesterase/lipase family.
  • Post-translational
    modifications

    Contains sialic acid.
    Cleavage of the signal sequence can occur at 2 positions, either between Trp-17 and Gly-18 or between Gly-18 and His-19.
  • Cellular localization

    Endoplasmic reticulum lumen.
  • Information by UniProt
  • Database links

  • Alternative names

    • ACAT antibody
    • Acyl coenzyme A cholesterol acyltransferase antibody
    • Acyl-coenzyme A:cholesterol acyltransferase antibody
    • Brain carboxylesterase hBr1 antibody
    • Carboxyesterase ES-3 antibody
    • Carboxylesterase antibody
    • Carboxylesterase 1 (monocyte/macrophage serine esterase 1) antibody
    • Carboxylesterase 1 antibody
    • Carboxylesterase 1 deficiency, included antibody
    • Carboxylesterase 2, formerly antibody
    • CE 1 antibody
    • CEH antibody
    • Ces-1 antibody
    • CES1 antibody
    • CES2 antibody
    • CESDD1 antibody
    • Cholesterol ester hydrolase, neutral, macrophage-derived antibody
    • Cholesteryl ester hydrolase antibody
    • Cocaine carboxylesterase antibody
    • EC 3.1.1.1 antibody
    • Egasyn antibody
    • ES-HTEL antibody
    • ES-x antibody
    • Es22 antibody
    • EST1_HUMAN antibody
    • Esterase 22 antibody
    • Esterase antibody
    • hCE 1 antibody
    • HMSE antibody
    • HMSE1 antibody
    • Liver carboxylesterase 1 antibody
    • Liver carboxylesterase 3 antibody
    • Methylumbelliferyl acetate deacetylase 1 antibody
    • MGC117365 antibody
    • MGC156521 antibody
    • Monocyte carboxylesterase deficiency, included antibody
    • Monocyte esterase deficiency, included antibody
    • Monocyte/macrophage serine esterase antibody
    • PCE-1 antibody
    • pI 5.5 esterase antibody
    • Proline-beta-naphthylamidase antibody
    • REH antibody
    • Retinyl ester hydrolase antibody
    • Serine esterase 1 antibody
    • Ses-1 antibody
    • SES1 antibody
    • TGH antibody
    • Triacylglycerol hydrolase antibody
    see all

Images

  • All lanes : Anti-Liver Carboxylesterase 1/CES1 antibody (ab45957) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole cell lysate
    Lane 2 : THP1 whole cell lysate (ab7913)
    Lane 3 : Liver (Rat) Tissue Lysate - normal tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 63 kDa

  • ICC/IF image of ab45957 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 (ab45957, 1µg/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). This antibody also gave a positive result in 100% Methanol fixed (5 min) HepG2 cells at 1µg/ml.
  • IHC image of Carboxylesterase 1 staining in Human normal liver 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 ab45957, 1µg/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:

  • Wen X  et al. Hepatic carboxylesterases are differentially regulated in PPARa-null mice treated with perfluorooctanoic acid. Toxicology 416:15-22 (2019). Read more (PubMed: 30685356) »
  • Xu J  et al. Global inactivation of carboxylesterase 1 (Ces1/Ces1g) protects against atherosclerosis in Ldlr-/-mice. Sci Rep 7:17845 (2017). Read more (PubMed: 29259301) »
See all 7 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question

In both experiments - how much protein are you loading? 25ug

Is it possible to increase this? As you can see my data, I did cellular fractionation and I expected that these two proteins show strong expression inmicrosomal fraction. It is known that AADACL1(or NCEH-1, KIAA1363) and liver carboxylesterase 1(or CES1) are expressed in microsome. According to my data, one was veryvery weak even I used a super signalECL for o/n and the other one was shown too much non-specific bands even in a positive control (THP-1).

Also, have you tested a shorter block or using BSA instead of milk? No, I didn't.

Are the antibodies diluted in block or PBS-T alone? In 5% skim milk

Regarding ab111544 - have you tried increasing the concentration of primary antibody?

No I didn't. But I think that is not the best solution because I got a very weak signal in spite of exposing a film with a super signal ECL for o/n. I think it will be no big differences if I increase the concentration of primary antibody.

Have you varied the concentration of secondary antibody to help optimize your results?

No I didn't. I can do troubleshooting most of cases. The secondary antibody concentration was 1:5,000. Do you think I need more less dilution of secondary antibody like 1:2,000? When I do western blot, lusually use 1:10,000 of secondary antibody. So I thought 1:5,000 was good starting dilution.(ab111544)

Similarly, with ab45957, have you tried altering the concentration of primary antibody used?

If I will western blot again with it, I will do dilute more. But I'm not sure this antibody is working well. Please check my data again. In an antibody data sheet, THP-1 was used a kind of positive controls. I also used THP-1 and differentiated THP-1 by PMA. But there were too many bands.

It seems that you are getting a very dark band at the predicted molecular weight, but further optimization is necessary. I would recommend decreasing the amount of primary and secondary antibodies to help clean up this blot.

I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

Thanks a lot for your time. I think I can test more in case of ab45957 at least. But in case of ab111544, I don't think so. I don't have much time for this. Please check my data again.

Thank you again, and have a nice day. Feel free to tell me if you need more information about my experiment.

Read More
Answer

Thank you for your reply.


Can you please provide an order number for your purchase of these antibodies? If you are contacting us within 6 months of purchase, I am happy to offer a replacement or credit per our Abpromise guarantee.


Regarding ab111544, are you looking to try another vial of this antibody or receive a credit? If you decide to test ab45957 again and are still having difficulties, I am happy to offer the same for this antibody.

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

Read More

Answer

Thank you for your reply with this protocol information.


I am sorry that you have been experiencing difficulties with two of our antibodies in WB. I have reviewed the protocol information you provided and I would like to ask some additional questions andmake some suggestions to improve your results.


In both experiments - how much protein are you loading? Is it possible to increase this? Also, have you tested a shorter block or using BSA instead of milk? Are the antibodies diluted in block or PBS-T alone?


Regarding ab111544 - have you tried increasing the concentration of primary antibody? Have you varied the concentration of secondary antibody to help optimize your results?


Similarly, with ab45957, have you tried altering the concentration of primary antibody used? It seems that you are getting a very dark band at the predicted molecular weight, but further optimization is necessary. I would recommend decreasing the amount of primary and secondary antibodies to help clean up this blot.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

Read More

Answer

Thank you for your message which has been forwarded to me as my colleague is currently away from the office.  I am sorry to hear you have had some difficulty detecting the recombinant protein. I can confirm that  often the presence of a flag tag can prevent access of an antibody to an epitope on recombinant proteins.  Particularly if the antibody is working on endogenous samples, this may be the case.  The immunogen for ab45957 Anti-Liver Carboxylesterase 1 antibody is from amino acids 548-567: TNLFAKKAVEKPPQTEHIEL I would also recommend to check the stability of the transfection and also the stability of the recombinant protein.  Sometimes fragments rather than the full recombinant protein could be expressed. I hope this information will be helpful to you.  Should you have any further questions, please do not hesitate to contact us.

Read More

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