Overview

  • Product name
    Anti-Apolipoprotein A I antibody
    See all Apolipoprotein A I primary antibodies
  • Description
    Goat polyclonal to Apolipoprotein A I
  • Host species
    Goat
  • Specificity
    Typically less than 1% cross reactivity against other types of apoLipoprotein was detected by ELISA. This antibody reacts with mouse apoLipoprotein A-I and has negligible cross-reactivity with Type A-II, B, C-I, C-II, C-III, E and J apoLipoproteins.
  • Tested applications
    Suitable for: Sandwich ELISA, ELISA, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Full length native apoLipoprotein Type A-I (purified).

  • General notes
    This antibody has been used to determine that atherosclerotic lesions in the human aorta contain considerable amounts of lipoproteins. These lipoproteins were observed to be complexed with components of the extracellular matrix (especially LDL and proteoglycans). The role of these matrix-lipoprotein complexes is not entirely clear, however, animal models of atherosclerosis have shown that increased cellular proliferation and increased production of extracellular matrix components occur following injury to the intimal layer of the aorta.

Properties

Applications

Our Abpromise guarantee covers the use of ab7614 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
Sandwich ELISA 1/5000 - 1/10000. Can be paired for Sandwich ELISA with Rabbit polyclonal to Apolipoprotein A I (ab20453).
ELISA 1/5000 - 1/10000.
IHC-P 1/250 - 1/500.
WB 1/500 - 1/1000.
IP 1/100.

Target

  • Function
    Participates in the reverse transport of cholesterol from tissues to the liver for excretion by promoting cholesterol efflux from tissues and by acting as a cofactor for the lecithin cholesterol acyltransferase (LCAT). As part of the SPAP complex, activates spermatozoa motility.
  • Tissue specificity
    Major protein of plasma HDL, also found in chylomicrons. Synthesized in the liver and small intestine.
  • Involvement in disease
    Defects in APOA1 are a cause of high density lipoprotein deficiency type 2 (HDLD2) [MIM:604091]; also known as familial hypoalphalipoproteinemia (FHA). Inheritance is autosomal dominant.
    Defects in APOA1 are a cause of the low HDL levels observed in high density lipoprotein deficiency type 1 (HDLD1) [MIM:205400]; also known as analphalipoproteinemia or Tangier disease (TGD). HDLD1 is a recessive disorder characterized by the absence of plasma HDL, accumulation of cholesteryl esters, premature coronary artery disease, hepatosplenomegaly, recurrent peripheral neuropathy and progressive muscle wasting and weakness. In HDLD1 patients, ApoA-I fails to associate with HDL probably because of the faulty conversion of pro-ApoA-I molecules into mature chains, either due to a defect in the converting enzyme activity or a specific structural defect in Tangier ApoA-I.
    Defects in APOA1 are the cause of amyloid polyneuropathy-nephropathy Iowa type (AMYLIOWA) [MIM:107680]; also known as amyloidosis van Allen type or familial amyloid polyneuropathy type III. AMYLIOWA is a hereditary generalized amyloidosis due to deposition of amyloid mainly constituted by apolipoprotein A1. The clinical picture is dominated by neuropathy in the early stages of the disease and nephropathy late in the course. Death is due in most cases to renal amyloidosis. Severe peptic ulcer disease can occurr in some and hearing loss is frequent. Cataracts is present in several, but vitreous opacities are not observed.
    Defects in APOA1 are a cause of amyloidosis type 8 (AMYL8) [MIM:105200]; also known as systemic non-neuropathic amyloidosis or Ostertag-type amyloidosis. AMYL8 is a hereditary generalized amyloidosis due to deposition of apolipoprotein A1, fibrinogen and lysozyme amyloids. Viscera are particularly affected. There is no involvement of the nervous system. Clinical features include renal amyloidosis resulting in nephrotic syndrome, arterial hypertension, hepatosplenomegaly, cholestasis, petechial skin rash.
  • Sequence similarities
    Belongs to the apolipoprotein A1/A4/E family.
  • Post-translational
    modifications
    Palmitoylated.
    Phosphorylation sites are present in the extracelllular medium.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apo-AI antibody
    • ApoA I antibody
    • ApoA-I antibody
    • APOA1 antibody
    • APOA1_HUMAN antibody
    • Apolipoprotein A-I(1-242) antibody
    • Apolipoprotein A1 antibody
    • Apolipoprotein AI antibody
    • Brp14 antibody
    • Ltw1 antibody
    • Lvtw1 antibody
    • Sep1 antibody
    • Sep2 antibody
    see all

Images

  • ab7614 staining Apolipoprotein A I in mouse liver tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before enzymatic antigen retrieval with Proteinase K and then blocking with 2% BSA for 20 minutes was performed. The primary antibody was diluted 1/6000 and incubated with sample for 24 hours at 4°C in 1% BSA. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/200 as secondary antibody.

    See Abreview

  • All lanes : Anti-Apolipoprotein A I antibody (ab7614) at 1/5000 dilution

    All lanes : Whole tissue lysate prepared from mouse liver

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : Donkey anti-goat Alexa Fluor® 647
    at 1/2500 dilution

    Observed band size: 25 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.



    Unknown band at 40kDa is well differentiated from the Apolipoprotein A I band at 25kDa.

References

This product has been referenced in:
  • Cao J  et al. Protein markers of dysfunctional HDL in scavenger receptor class B type I deficient mice. J Transl Med 16:155 (2018). WB ; Mouse . Read more (PubMed: 29879989) »
  • Cash JG & Hui DY Liver-specific overexpression of LPCAT3 reduces postprandial hyperglycemia and improves lipoprotein metabolic profile in mice. Nutr Diabetes 6:e206 (2016). Read more (PubMed: 27110687) »
See all 10 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
10 µg
Specification
Liver
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-20 %)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Oct 02 2012

Answer

Thank you for contacting us. We currently only have a mouse ELISA kit for Apolipoprotein CIII: ab108811 However, we do have antibodies to each of your targets of interest that can be used in ELISA on mouse samples: Apolipoprotein A I: ab7614 or ab20453 Apolipoprotein B: ab117317 Apolipoprotein CIII: ab55984 If you could let me know which products you are interested in and what quantities, I would be happy to create a proforma invoice for you.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver, stomach, kidney, testes, lung, heart, etc)
Specification
Liver, stomach, kidney, testes, lung, heart, etc
Fixative
Formaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Proteinase K
Permeabilization
No
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 2%

Miss. Judit Marsillach

Verified customer

Submitted Jul 24 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Loading amount
50 µg
Specification
liver
Gel Running Conditions
Reduced Denaturing (9%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Hua Jiang

Verified customer

Submitted Jan 02 2009

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Purified protein (purified protein (BIODESIGN))
Loading amount
0.5 µg
Specification
purified protein (BIODESIGN)
Gel Running Conditions
Reduced Denaturing
Blocking step
5%mill+1%BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 11 2008

Answer

Thank you for your enquiry. This product was tested using a direct ELISA using purified mouse Apo A-1. It was not tested using different concentrations of mouse serum. I am sorry you are experiencing problems with this antibody. Below please find a link to our troubleshooting questionnaire. This will give us further information regarding the samples you tested and the protocol you used. Once you have submitted the form to us, we will look at the protocol you used and see if there are any suggestions we can make to improve your results. The questionnaire can be found at: https://www.abcam.com/index.html?section=elisa&pageconfig=technical&intAbID=7614&mode=questionaire I look forward to receiving your reply.

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Question

BATCH NUMBER 57140 ORDER NUMBER 58914 DESCRIPTION OF THE PROBLEM The antibody did not stain specific protein in arterial tissue (immunhistochemistry of formalin fixed tissue), in peripheral blood mononuclear cells (cytospin of native cells) nor in plasma (western blot). The only detectable signal in the tissue was in leukocytes (presumably false positive peroxidase signal). Simultaneously we used your antibody against ApoB100 (ab7616) which worked perfectly fine, both in tissue and in the western blot. SAMPLE human arterial tissue, liver and kidney. human plasma human peripheral blood mononuclear cells (PBMC) PRIMARY ANTIBODY tissue: ab7614; 10?g/ml (1:100; 1: 50 and 1:200 were also tested) in Antibody diluent (DAKO S2022); 1h RT Western blot: ab7614; 1?g/ml 5% fat free milk/TBS, 1h RT wash: tissue 2x in PBS; Western blot 3x in TBS. SECONDARY ANTIBODY tissue: Medite, NCL-RTU-Qu. Novostain Universal Quick kit, 30min RT Western blot: peroxidase conjugated rabbit anti goat DAKO (P0449) 0.1?g/ml in 5% fat-free milk/TBS; 1h RT wash: tissue 2x in PBS; Western blot 3x in TBS. DETECTION METHOD tissue: DAB (DAKO K3468) western blot: super signal West Pico kit (Pierce 1856135 + 1856136) POSITIVE AND NEGATIVE CONTROLS USED Plasma contains known amount of ApoA1 (1.5mg/ml) as a positive control tissue we used liver. we did not use purified protein nor deficiency plasma. ANTIBODY STORAGE CONDITIONS after arrival, 5 aliquots (20?l) were pipeted and stored frozen at -20? after thawing one aliquot for usage, the ab stored at 4? and not refrozen. the interval between antibody arrival and aliquotation was 5 days and the first aliquot was immediately. FIXATION OF SAMPLE tissues were fixed in 4% phosphate buffered formalin plasma and PBMC were used unfixed ANTIGEN RETRIEVAL used only for formalin fixed tissues: proteinase K was used according to the manufacturer's description (DAKO S3020) alternatively we tried citrate buffer in heat pressure chamber but this resulted in high background staining PERMEABILIZATION STEP The only procedure were permeabilization was used was in cytospin preps of PBMCs. We used 0.1% saponin in PBS-FCS. BLOCKING CONDITIONS for tissue: Medite, normal horse serum, NCL-H-serum, ready for use for western blot: 5% fat free milk in TBS (pH 7.4) for cytospin: 1% BSA-1%FCS in PBS (pH 7.4) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary antibody dilution alternative detection approaches such as western blot. different presumably positive tissues. more 300 arterial specimens were stained and were negative ! we ordered a second antibody lot ! ADDITIONAL NOTES Since we are very interested in the reliable detection of apolipoprotein A1 in arterial tissues we are looking forward to your suggestions.

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Answer

Thank you for your enquiry and for taking your time to fill in the on-line Questionnaire. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. We would like to emphasize the fact that this antibody has been tested for cross-reaction only with mouse but other species have not been characterized yet. Therefore, unfortunately, we do not know if it recognizes human Apolipoprotein A I. We would suggest using mouse samples as positive control along with the human samples to make sure that the detection system works properly. Since you are using this antibody on human samples which has not been tested yet, we would appreciate any feedback information about your results which we could share with our present and future customers. Please let us know how you get on, and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on a number of rewards. We hope this information will be useful for you.

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Answer

Thank you for your enquiry. We would suggest performing a regular sandwich ELISA protocol.If you need anything further or any help then please let us know.

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Answer

Thank you for your enquiry. The immunogen is from mouse that's why the antibody recognizes mouse Apolipoprotein A I.

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Answer

Thank you for your enquiry. Unfortunately, the purified Apolipoprotein A I protein which has been used is not commercially available.

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1-10 of 12 Abreviews or Q&A

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