Overview

  • Product name

  • Description

    Rabbit polyclonal to AADACL1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Fr, ICCmore details
  • Species reactivity

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

    Synthetic peptide corresponding to a region within internal amino acids 224-273 (LQALDFNTPS YQQSMNTPIL PRHVMVRYWL DYFKGNYDFV EAMIVNNHTS) of Mouse AADACL1 (NP_848887).

  • Positive control

    • Mouse kidney lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab111544 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/IF Use at an assay dependent concentration.
WB Use a concentration of 0.2 - 1 µg/ml. Predicted molecular weight: 46 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-Fr Use at an assay dependent concentration.
ICC 1/200.

Target

  • Function

    Hydrolyzes 2-acetyl monoalkylglycerol ether, the penultimate precursor of the pathway for de novo synthesis of platelet-activating factor. May be responsible for cholesterol ester hydrolysis in macrophages, thereby contributing to the development of atherosclerosis. Also involved in organ detoxification by hydrolyzing exogenous organophosphorus compounds. May contribute to cancer pathogenesis by promoting tumor cell migration.
  • Tissue specificity

    Expressed in monocyte-derived macrophages. Up-regulated in invasive melanoma and breast carcinoma cell lines.
  • Sequence similarities

    Belongs to the 'GDXG' lipolytic enzyme family.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Membrane. Microsome.
  • Information by UniProt
  • Database links

  • Alternative names

    • Arylacetamide deacetylase like 1 antibody
    • Arylacetamide deacetylase-like 1 antibody
    • NCEH antibody
    • Nceh1 antibody
    • NCEH1_HUMAN antibody
    • Neutral cholesterol ester hydrolase 1 antibody
    • Neutral cholesterol ester hydrolase antibody
    see all

Images

  • Anti-AADACL1 antibody (ab111544) at 1 µg/ml + Mouse kidney lysate at 10 µg

    Predicted band size: 46 kDa



    Gel concentration: 12%
  • ab114544 staining AADACL1 in cultured human prostate cells by ICC (Immunocytochemistry). Cells were fixed with methanol and blocked with 1% BSA for 24 hours at 4°C. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 37°C. A Biotin-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab111544 staining AADACL1 in Human Prostate cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 1% BSA + 2% serum for 24 hours at 4°C. Samples were incubated with primary antibody (1/200 in blocking agent) for 2 hours at 37°C. An undiluted FITC-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

References

ab111544 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-7 of 7 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 contacting Abcam regarding ab111544.


I am sorry that you have been experiencing difficulties with this antibody in WB. In order to assist you further, would you please provide some additional details? Specifically a description of the sample and preparation as well as a summary of the WB protocol you followed (including incubation times and temperatures, dilutions, etc). Once I have this information, I would be happy to help troubleshoot or provide a replacement per our Abpromise guarantee.


I look forward to your reply so that I may assist you further.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Prostate tumour)
Specification
Prostate tumour
Fixative
Methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 10 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Prostate)
Specification
Prostate
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jun 08 2012

Application
Immunocytochemistry
Sample
Human Cultured Cells (Prostate)
Specification
Prostate
Fixative
Methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Dr. Olayinka Oloko

Verified customer

Submitted Apr 09 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (prostate)
Loading amount
25 µg
Specification
prostate
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Olayinka Oloko

Verified customer

Submitted Dec 07 2011

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