Overview

  • Product name
  • Description
    Goat polyclonal to CPT1A
  • Host species
    Goat
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Rhesus monkey
  • Immunogen

    Synthetic peptide:

    C-DPAQTVEQRLKLFK

    , corresponding to internal sequence amino acids 621-634 of Human CPT1A

  • Positive control
    • Human Liver lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab53532 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 - 3 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 86 kDa).

Target

  • Tissue specificity
    Strong expression in kidney and heart, and lower in liver and skeletal muscle.
  • Pathway
    Lipid metabolism; fatty acid beta-oxidation.
  • Involvement in disease
    Defects in CPT1A are the cause of carnitine palmitoyltransferase 1A deficiency (CPT1AD) [MIM:255120]; also known as CPT-I deficiency or CPT1A deficiency. CPT1AD is a rare autosomal recessive metabolic disorder of long-chain fatty acid oxidation characterized by severe episodes of hypoketotic hypoglycemia usually occurring after fasting or illness. Onset is in infancy or early childhood.
  • Sequence similarities
    Belongs to the carnitine/choline acetyltransferase family.
  • Cellular localization
    Mitochondrion outer membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Carnitine O palmitoyltransferase 1 liver isoform antibody
    • Carnitine O palmitoyltransferase I antibody
    • Carnitine O palmitoyltransferase I liver isoform antibody
    • Carnitine O-palmitoyltransferase 1 antibody
    • Carnitine O-palmitoyltransferase I antibody
    • Carnitine palmitoyltransferase 1A (liver) antibody
    • Carnitine palmitoyltransferase 1A antibody
    • Carnitine palmitoyltransferase I antibody
    • Carnitine palmitoyltransferase I liver antibody
    • CPT 1 antibody
    • CPT I antibody
    • CPT1 antibody
    • CPT1 L antibody
    • CPT1-L antibody
    • Cpt1a antibody
    • CPT1A_HUMAN antibody
    • CPTI antibody
    • CPTI-L antibody
    • L CPT1 antibody
    • liver isoform antibody
    see all

Images

  • ab53532 antibody (0.1µg/ml) staining of CPT1A in Human Liver lysate (35µg protein in RIPA buffer).
    Predicted band size: 86 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    Primary incubation was 1 hour. Detected by chemiluminescence.

References

This product has been referenced in:
  • Mazibuko-Mbeje SE  et al. Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30717198) »
  • Samudio I  et al. Pharmacologic inhibition of fatty acid oxidation sensitizes human leukemia cells to apoptosis induction. J Clin Invest 120:142-56 (2010). WB ; Human . Read more (PubMed: 20038799) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question

I will attempt to answer your questions below:
1. What's the species you were using to detect the proteins: human,
mouse, etc. proteins with a tag? HEK-293 lysates (human clones with
N-terminal V5-tag). Some abs from abcam (ab60332, ab80464) worked well.
2. Almost all of our antibodies are developed to work under denatured
conditions or otherwise it will be stated on the datasheet. It may be
that the antibody cannot detect its epitope as it may be simply not
accessible. Please boil your samples to provide optimal conditions.
The same goes for the reducing conditions of the gel which are
usually necessary for the most our antibodies to work properly.
I have tried denatured conditions are results were the same!
3. In order to avoid over-loading and therefore to decrease the side
bands, I would like to suggest loading 20 to 30 ug protein per lane.
I have loaded 20-40 ug protein per lane.
3. What are the listed primaries antibodies concentration? Or do you
use 1:5000 for all of them?
As you know, the primary antibody concentranctions vary, for instance:
ab15575 (1:5000), ab53532 (1:1000), ab96615 (1:1000) and ab106460 (1:1000)
4. What kind of secondary antibodies are you using for the listed
primaries? Do you perform a non-primary control with them beforehand?
depending on primary antibody used, secondary antibodies were: goat
anti-mouse IgGHRP (170-6516), goat anti-rabbit IgGHRP (170-6515) and
rabbit anti-goat IgGHRP (172-1034) from biorad.
5. Is it possible to send us an image?
6. How did you store the antibodies?
antibodies are stored as recommended, usually -20 C or sometimes 4C
P.s. note that I have already tested these antibodies at least 3 times
over the last 4 months. Hope to get replacement aliquots shortly.

Read More
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 I have mentioned before, this two orders are way out of our Abpromise. However, I can see from the details you kindly provided the antibodies should have worked under these conditions, and I am therefore willing to make an exception in this case.


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.

Read More

Question

Please find the WB questionnaire below:
1) Abcam product code ab: (ab15575, ab53532, ab96615 and ab106460)

3) Description of the problem: the mentioned antibodies are
non-specific i.e. several bands appear, none of which is the right size.
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…):
cytoplasmic extract
Lysis buffer : Lysis buffer (50 mM HEPES, pH 7.4, 150 mM NaCl, 1 mM
EDTA, 1% Nonidet P-40, 1 mM DTT and 10% glycerol) supplemented with
benzonase nuclease (250U, E1014, Sigma),
Protease inhibitors: 1 mM phenylmethylsulfonyl fluoride (P7626, Sigma)
and 1X protease inhibitor cocktail (Cat. No. 04693116001, Roche
Applied Science),
Phosphatase inhibitors
Reducing agent: 1 mM DTT
Boiling for ≥5 min? no
Protein loaded ug/lane or cells/lane: 60 ug
Positive control: V5-tagged constructs were detected with anti-V5 antibody
Negative control: empty cells
5) Percentage of gel: 10%
Type of membrane : nitrocellulose membrane
Protein transfer verified: by ponceau stain
Blocking agent and concentration: 5% Milk in TBS + 0.1% Tween 20
Blocking time: 2 h
Blocking temperature : RT
6) Was a fresh membrane used or had it been probed and stripped with a
different antibody? Yes
7) Primary antibody (If more than one was used, describe in
“additional notes”) : Mouse monoclonal anti-V5 (R960-25, Invitrogen)
Concentration or dilution: 1:5,000
Diluent buffer : 3% BSA in TBS
Incubation time: 16 h
Incubation temperature: 8 C
8) Secondary antibody: goat anti-mouse IgGHRP (biorad# 170-6516)
Species:
Reacts against: mouse
Concentration or dilution: 1:25,000
Diluent buffer : 3% BSA in TBS
Incubation time: 45 min
Incubation temperature: RT
Fluorochrome or enzyme conjugate:IgG-HRP
9) Washing after primary and secondary antibodies:
Buffer
Number of washes: 2
10)Detection method: autoradiography
11) How many times have you run this staining? 3
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody?
the anti-V5 antibody used has already been tested in the lab and found
to work well.
BR
Enzo
--
Enzo Scifo, PhD candidate
Meilahti Clinical Proteomics Core Facility
Institute of Biomedicine/Anatomy
P.O.Box 63 (Haartmaninkatu 8)
FI-00014 University of Helsinki
Helsinki, Finland
Tel: +358919125202
Fax: +358919125206
Mobile: +358466370309

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I can confirm that unfortunately, as you purchased this product farover 180 days ago it is will no longer be covered by our Abpromise guarantee on this occasion. Details of our guarantee are on our website and we encourage customers to contact us as soon as possible if they experience any difficulties.If a particular antibody causes problems we do encourage customers to contact us as soon as possible to save time and material. Although not covered by our guarantee, we would still be pleased to offer some suggestions to help optimize the results

Having reviewed this case, I would like to offer some suggestions to help optimize the resultsandI would also appreciate if you can confirm some further details:

1. What's the species you were using to detect the proteins: human, mouse,etc. proteins with a tag?

2. Almost all of our antibodies are developed to work under denatured conditions or otherwise it will be stated on the datasheet. It may be that the antibody cannot detect itsepitope as it may be simply not accessible. Please boil your samples to provide optimal conditions. The same goes for the reducing conditions of the gel which are usually necessary forthe most our antibodies to work properly.

3.In order to avoid over-loading and therefore to decrease the side bands, I would like to suggest loading 20 to 30 ug protein per lane.

3. What arethe listedprimaries antibodies concentration?Or do you use 1:5000 for all of them?

4. What kind of secondary antibodies are you using for the listed primaries? Do you perform a non-primary control with them beforehand?

5. Is it possible to send us an image?

6. How did you store the antibodies?


Should the suggestions not improve the results, please do let me know.


I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.


If you wouldn't mind, I would like to try to understand what may be contributing to the problems encountered in order to hopefully resolve the situation. To this end, could you please fill in the questionnaire I have attached to this email. If you could include images of the results obtained (including that using the V5 antibody) that would be very helpful.

I also have a few additional questions:

1. How were the V5-protein construct made? Which sequences were used to construct the fusion protein (human/mouse/rat etc)?

2. Which V5 antibody was used? And which secondary antibody was used with this antibody?

3. From your orders I can see that you also purchased ab80464, ab60332 and ab84589, were you using the same protocol using these (V5 tagging?)? Did you have any problems in using these antibodies as well?

OnceIhavereceive this information and the completed questionnaire,I will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.

I look forward to receiving your reply.

Read More

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