Overview

  • Product name
  • Description
    Rabbit polyclonal to DGAT1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Zebrafish
  • Immunogen

    A synthetic peptide designed within residues 200-300 of human DGAT1.

  • Positive control
    • HepG2 lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab54037 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 2 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 23499441

Target

  • Function
    Catalyzes the terminal and only committed step in triacylglycerol synthesis by using diacylglycerol and fatty acyl CoA as substrates. In contrast to DGAT2 it is not essential for survival. May be involved in VLDL (very low density lipoprotein) assembly.
  • Pathway
    Lipid metabolism; glycerolipid metabolism.
  • Sequence similarities
    Belongs to the membrane-bound acyltransferase family. Sterol o-acyltransferase subfamily.
  • Cellular localization
    Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACAT related gene product 1 antibody
    • ACAT-related gene product 1 antibody
    • Acyl coenzyme A:cholesterol acyltransferase related gene 1 antibody
    • Acyl-CoA retinol O-fatty-acyltransferase antibody
    • Acyl-CoA:diacylglycerol acyltransferase antibody
    • ARAT antibody
    • ARGP1 antibody
    • C75990 antibody
    • D15Ertd23e antibody
    • Dgat antibody
    • DGAT1 antibody
    • DGAT1_HUMAN antibody
    • Diacylglycerol O acyltransferase 1 antibody
    • Diacylglycerol O-acyltransferase 1 antibody
    • DIAR7 antibody
    • Diglyceride acyltransferase antibody
    • EC 2.3.1.20 antibody
    • hCG_24006 antibody
    • MGC139064 antibody
    • Retinol O fatty acyltransferase antibody
    see all

Images

  • Anti-DGAT1 antibody (ab54037) at 2 µg/ml + HepG2 lysate

    Predicted band size: 55 kDa
    Observed band size: 55 kDa

References

This product has been referenced in:
  • Angeli E  et al. Protein and gene expression of relevant enzymes and nuclear receptor of hepatic lipid metabolism in grazing dairy cattle during the transition period. Res Vet Sci 123:223-231 (2019). Read more (PubMed: 30684909) »
  • Feng S  et al. Proton pump inhibitor pantoprazole inhibits the proliferation, self-renewal and chemoresistance of gastric cancer stem cells via the EMT/ß-catenin pathways. Oncol Rep 36:3207-3214 (2016). Read more (PubMed: 27748935) »
See all 9 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1278065.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thank you for contacting us.

The sequence homology between human and Drosophila is only 42% and the immunogen sequence homology is also very less for cross reactivity. I am sorry to say this antibody might not be suitable for Drosophila species.

If you however would like to try this antibody. Then please contact me for Abtrial discount code. Click the link for more details about Abtrial;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for your reply and the sequence.

Unfortunately, after investigation I confirm that none of the 4 anti-DGAT1 antibodies is likely to work in Drosophila. The immunogens used to raise ab59034, ab122924 and ab100982 do not show more than 46% homology with the fly DGAT1.
I would like to recommend checking the Biocompare website which has an excellent antibody search facility that includes many suppliers. The link is http://www.biocompare.com.

We are happy to be able to offer custom antibody services through Epitomics, an Abcam company. For more information about these services visit http://www.epitomics.com/services/ or email mailto:service@epitomics.com.
Alternatively, please contact Li Li at Epitomics: mailto:li.li@epitomics.com

I hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions in this regard.

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Answer

Thank you for contacting us.

Unfortunately, the source of the antibody informed us that Drosophila and human DGAT1 sequences are too divergent to recommend ab54037 for the detection of DGAT1 in fly.

We have 3 other references that may be suitable for your work. In order to determine if one of these alternative antibodies would be suitable for fruit fly, could you please send me the full length sequence of the fly DGAT1?

Thank you.

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Answer

Thank you for your e-mail and for attaching the scanned Western blot image.

My colleague is out of office for a couple of days this week and she has asked me to look after her customers during her absence.

The band appears to be at around 26 kDa on the picture. It could be some degradation product though as my colleague has explained to you in her previous e-mail, there is some published reference suggesting that protease activity can reveal a fragment of 25 kDa.

http://www.jbc.org/content/285/48/37377.full
Expand+Journal of Biological Chemistrywww.jbc.orgFirst Published on September 27, 2010, doi: 10.1074/jbc.M110.163691 November 26, 2010 The Journal of Biological Chemistry, 285, 37377-37387. Topological Orientation of Acyl-CoA:Diacylglycerol Acyltransferase-1 (DGAT1) and Identification of a Putative Active Site Histidine and the Role of the N Terminus in Dimer/Tetramer Formation Pamela J. McFie et al

In order to be able to hl you further in this case, it would be very useful for us to see the protocol used in the experiments. We are particularly interested in the followings:

- Lysis buffer used during sample preparation:

- Loading control:

- Positive control:

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWBand mouse. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

Reviewing the details, I have checkedthe UniProt protein database entry for the DGAT1 protein anddone a short literature search whichdo not indicate there areany isoforms of splice varients. However, I have found the following references which indicates that protease activity can reveal a fragment of 25 kDa which may be helpful. I can suggest to considera further literature search to obtain more information.

http://www.jbc.org/content/285/48/37377.full
Expand+Journal of Biological Chemistrywww.jbc.orgFirst Published on September 27, 2010, doi: 10.1074/jbc.M110.163691 November 26, 2010 The Journal of Biological Chemistry, 285, 37377-37387.
Topological Orientation of Acyl-CoA:Diacylglycerol Acyltransferase-1 (DGAT1) and Identification of a Putative Active Site Histidine and the Role of the N Terminus in Dimer/Tetramer Formation
Pamela J. McFie et al

Ensuring the sample is fresh and that protease inhibitors are included with the lysis buffer may help optimize the results in this case.

If you still have concerns regarding the results, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could also provide an image, including molecular weight markers,which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

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Question

LOT NUMBER GR27821-3 ORDER NUMBER 990441 DESCRIPTION OF THE PROBLEM Non-specific band SAMPLE HepG, Huh cell PRIMARY ANTIBODY 1:200 DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Yes ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Lysis buffer: 50 mM Tris-HCl (pH 8), 150 mM NaCl, 1% Triton X-100 Protease inhibitors: Roche protease inhibitor cocktail tablets (# 11 697 498 001) Boiling for ≥5 min? yes =boiling for 5 min Protein loaded ug/lane or cells/lane 50 - 100 ug/lane Positive control : HepG2 cell lysates Negative control : 293T cell lysates AMOUNT OF PROTEIN LOADED 50 - 100 ug/lane ELECTROPHORESIS/GEL CONDITIONS 8% gel 0.5 BSA in PBS overnight 4C TRANSFER AND BLOCKING CONDITIONS PVDF membrane, 5% skim milk, SECONDARY ANTIBODY goat anti rabbit HRP conjugate PBST, 3times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Dilution factor ADDITIONAL NOTES The purpose of our experiment is to detect DGAT-1 protein in Huh cell line. According to data sheet from your company, we used HepG2 cell line as a positive control, and 293T cell line as a negative control. And then because we could not detect DGAT-1 in Huh cells lysate, we again purchased DGAT-1 antibody from Santa-Cruz biotech. Using santa-cruz Ab, we could detect DGAT-1 in HepG2 cells lysate as a positive control, could not detect in 293T cells lysate as a negative control. So, we suggest that your DGAT-1 Ab is not working for our experiment, and want to replace with another DGAT-1 or any other Ab that we required.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will provide us with vital information for our monitoring of product quality I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation. Although there are no major tips to provide, it may be beneficial to consider the following for future experiments: 1. We recommend to load 20 - 30 ug of protein per lane of the gel to ensure it is not overloaded. 2. I can suggest to try BSA rather than milk to block. Changing blocking agent can sometimes help to improve results. 3. Was a loading buffer used, such as Laemmli buffer containing SDS and mercaptoethanol to reduce and denature? Thiscan be added to the lysate before boiling for 5 minutes. This will ensure the proteins are in the correct conformation to run at the correct molecular weight and be detected by the antibody. 4. I can suggest to consider trying the antibody at a lower concentration of 1:1000 to help reduce the background. 5. Is the current vial of secondary antibody working well with other primary antibodies? I can suggest it would be beneficial to consider including a no primary control to assess if the secondary antibody is binding non specifically. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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