Overview

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

    Full length native protein (purified)(Human) (isolated from human plasma by density gradient centrifugation followed by HPLC purification).

  • Positive control
    • purified apolipoprotein E This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7.
  • 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 ab7620 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
Indirect ELISA 1/4000 - 1/8000.
WB 1/5000 - 1/10000. Predicted molecular weight: 36 kDa.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function
    Mediates the binding, internalization, and catabolism of lipoprotein particles. It can serve as a ligand for the LDL (apo B/E) receptor and for the specific apo-E receptor (chylomicron remnant) of hepatic tissues.
  • Tissue specificity
    Occurs in all lipoprotein fractions in plasma. It constitutes 10-20% of very low density lipoproteins (VLDL) and 1-2% of high density lipoproteins (HDL). APOE is produced in most organs. Significant quantities are produced in liver, brain, spleen, lung, adrenal, ovary, kidney and muscle.
  • Involvement in disease
    Defects in APOE are a cause of hyperlipoproteinemia type 3 (HLPP3) [MIM:107741]; also known as familial dysbetalipoproteinemia. Individuals with HLPP3 are clinically characterized by xanthomas, yellowish lipid deposits in the palmar crease, or less specific on tendons and on elbows. The disorder rarely manifests before the third decade in men. In women, it is usually expressed only after the menopause. The vast majority of the patients are homozygous for APOE*2 alleles. More severe cases of HLPP3 have also been observed in individuals heterozygous for rare APOE variants. The influence of APOE on lipid levels is often suggested to have major implications for the risk of coronary artery disease (CAD). Individuals carrying the common APOE*4 variant are at higher risk of CAD.
    Genetic variations in APOE are associated with Alzheimer disease type 2 (AD2) [MIM:104310]. It is a late-onset neurodegenerative disorder characterized by progressive dementia, loss of cognitive abilities, and deposition of fibrillar amyloid proteins as intraneuronal neurofibrillary tangles, extracellular amyloid plaques and vascular amyloid deposits. The major constituent of these plaques is the neurotoxic amyloid-beta-APP 40-42 peptide (s), derived proteolytically from the transmembrane precursor protein APP by sequential secretase processing. The cytotoxic C-terminal fragments (CTFs) and the caspase-cleaved products such as C31 derived from APP, are also implicated in neuronal death. Note=The APOE*4 allele is genetically associated with the common late onset familial and sporadic forms of Alzheimer disease. Risk for AD increased from 20% to 90% and mean age at onset decreased from 84 to 68 years with increasing number of APOE*4 alleles in 42 families with late onset AD. Thus APOE*4 gene dose is a major risk factor for late onset AD and, in these families, homozygosity for APOE*4 was virtually sufficient to cause AD by age 80. The mechanism by which APOE*4 participates in pathogenesis is not known.
    Defects in APOE are a cause of sea-blue histiocyte disease (SBHD) [MIM:269600]; also known as sea-blue histiocytosis. This disorder is characterized by splenomegaly, mild thrombocytopenia and, in the bone marrow, numerous histiocytes containing cytoplasmic granules which stain bright blue with the usual hematologic stains. The syndrome is the consequence of an inherited metabolic defect analogous to Gaucher disease and other sphingolipidoses.
    Defects in APOE are a cause of lipoprotein glomerulopathy (LPG) [MIM:611771]. LPG is an uncommon kidney disease characterized by proteinuria, progressive kidney failure, and distinctive lipoprotein thrombi in glomerular capillaries. It mainly affects people of Japanese and Chinese origin. The disorder has rarely been described in Caucasians.
  • Sequence similarities
    Belongs to the apolipoprotein A1/A4/E family.
  • Post-translational
    modifications
    Synthesized with the sialic acid attached by O-glycosidic linkage and is subsequently desialylated in plasma. O-glycosylated with core 1 or possibly core 8 glycans. Thr-307 is a minor glycosylation site compared to Ser-308.
    Glycated in plasma VLDL of normal subjects, and of hyperglycemic diabetic patients at a higher level (2-3 fold).
    Phosphorylation sites are present in the extracelllular medium.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • AD2 antibody
    • Apo-E antibody
    • APOE antibody
    • APOE_HUMAN antibody
    • APOEA antibody
    • Apolipoprotein E antibody
    • Apolipoprotein E3 antibody
    • ApolipoproteinE antibody
    • Apoprotein antibody
    • LDLCQ5 antibody
    • LPG antibody
    see all

Images

  • Anti-Apolipoprotein E antibody (ab7620) at 1/1000 dilution (in PBS/Tween (0.01%) + 3% milk for 12 hours) + Whole cell lysate of Rat glial cells at 40 µg

    Secondary
    An HRP-conjugated Rabbit anti-goat Polyclonal at 1/200 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 35-36 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.



    Blocking Step: 3% Milk for 20 minutes at room temperature

    See Abreview

  • ICC/IF image of ab7620 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey 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 ab7620 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- goat (ab96931) IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Minghui Z  et al. Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics. Proteomics Clin Appl N/A:e1700153 (2018). Read more (PubMed: 29999587) »
  • Fan J  et al. Identification of a Chrysanthemic Ester as an Apolipoprotein E Inducer in Astrocytes. PLoS One 11:e0162384 (2016). ELISA ; Human . Read more (PubMed: 27598782) »
See all 10 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - other (s2 cells)
Specification
s2 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 30 2013

Answer

Thank you for contacting us and your interest in our products.

Unfortunately, none of the three antibodies you are interested in have been tested in in both the application you are interested in with rat samples and we do not have any other antibodies against Apolipoprotein E which have either.

Ab7620 has been shown to detect the rat Apolipoprotein E, but has not been used in IHC. Ab1906 has been used in IHC with paraffin embedded sections but not with tissue from rat. From the sequence homology of the immunogen used I would also suggest that this anitbody would be unlikely to cross react with protein from rat. Finally, ab20874 has been used with rat samples and IHC with frozen sections, but not in paraffin embedded sections. We cannot therefore guarantee how any of these antibodies would perform in your experiment. I would suggest that ab23832 may be the most suitable to try to see if it would be suitable for your experiment.

We do however have an Abtrial program which may be of interest to you. This offer means that if you purchase ab23832 as normal, test the antibody with your rat samples in IHC-P and let us know of the results through an Abreview (no matter whether positive or negative) you would then be eligible to claim the value of ab23832 off any product in our catalogue. More information on this offer can be found here:

www.abcam.com/abtrial

Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab23832.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

Read More

Answer

Thank you for your reply.


I am happy to hear that this antibody worked well for you in IHC-P. Please be advised that the code that was issued to you is a credit note, since the information on our website was found to be incorrect. It can be used at any time and towards any invoice.


This is separate from our testing discount option. For testing discounts, customers must contact us prior to purchasing a product to determine if a product is eligible for the discount. Once this is determined, a code is issued. In order to redeem the code, you must purchase the antibody you wish to test, test it in an untested application and/or species, and submit an Abreview of your results. Once the review is submitted, the code can be redeemed for a free primary antibody on a future order.


Please visit our website for additional details: https://www.abcam.com/collaborationdiscount.


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

Read More

Question
Answer

Thank you for your inquiry.


I heard back from the supplying laboratory that this antibody has not, in fact, been tested in IHC-P. Data for IHC-P for their anti-ApoA2 antibody (which is produced to similar specifications) suggests the anti-ApoE antibody will work well for IHC using suitable samples and conditions, however the ApoE antibody has not been validated in IHC-P. I have since removed this application from the list on the datasheet, and I sincerely apologize for the error.


Since ab7620 hasn'tbeen validated in IHC-P,I can offer a discount off a future purchase if you buy ab7620 now, test it in IHC-P and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab7620 in IHC-P. I will then send a discount code. This code must be issued before purchasingab7620 so please wait for my reply before ordering.

2. Purchase ab7620 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab7620 turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (glial cells)
Loading amount
40 µg
Specification
glial cells
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: RT°C

Dr. GOSSELET Fabien

Verified customer

Submitted Nov 30 2010

Question

BATCH NUMBER 17739 ORDER NUMBER ab7620 DESCRIPTION OF THE PROBLEM wrong band size at 85 kDa SAMPLE plasma derived enriched chylomicrons, ldl, hdl and plasma PRIMARY ANTIBODY ApoE (Abcam #ab7620 as above) 1:5000 in PBS with 0.3% Tween with 2% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)for 1h, three times washing with PBS with 0,3% Tween 20 SECONDARY ANTIBODY Donkey Anti-Goat IgG (H+L) (Jackson; Cat# 705-036-147)1:20000 in PBS with 0.3% Tween with 2% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)for 1h, three times washing with PBS with 0,3% Tween 20 DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED plasma derived enriched chylomicrons, ldl, hdl ANTIBODY STORAGE CONDITIONS -20?C SAMPLE PREPARATION precipitated proteins resolved in laemmli buffer (according to Jouranl of Lipi Research Volume 30, 1989 S757-761 (Maguire GF, Lee M, Connelly PW)Sodium dodecyl sulfate-glycerol polyacrylamide slab gel electrophoresis for the resolution of apolipoproteins. AMOUNT OF PROTEIN LOADED 27 ?g protein ELECTROPHORESIS/GEL CONDITIONS reducing (mercaptoethanol),12% SDS PAGE Gel TRANSFER AND BLOCKING CONDITIONS 25mM Tris; 190mM Glycin; 1mM EDTA; 0.05% SDS; 20% Ethanol, time 1h at 60 V; 5% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)in PBS with 0.3% Tween 20 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? the first dilution of the primary antibody was 1:1000 which was too less diluted because we have multiple unspecific bands and with 1:10000 dilution the signal was to weak there was only a signal at 85 kDa this unspefic band at 85 kDa we still have at a antibody dilution of 1:5000 which we used in the later experiments

Read More
Answer

I'm sorry to hear you are having problem with ab7620. This is the first complaint we receive about this popular antibody. We would recommend using this antibody at a 1:5000 to 1:10 000 dilution, and I am puzzled at why you detect an 85kDa band. Having researched on what possible reasons this might be, I have found out that the protein has several isoforms, a potential carbohydrate chain can be added onto some forms, and it has several binding sites: for heparin (5-30kDa), beta amyloid as well as the receptors VLDL (160kDa) and apoER2 (10.2kDa). I have contacted the source of this antibody to ask for suggestions and also for the protocol used to test the antibody. Unfortunately there may be a delay of more than a week as the person is currently away. My sincere apologies for the delay,

Read More

Answer

Thank you for your enquiry. The antibody was made against whole apo E. It should react with all isoforms which enrich in Human plasma. If you have any further questions please do not hesitate to get in touch.

Read More

Answer

Thank you for your enquiry. Ab7620 has been tested with cross-reactivity with human and has not yet been tested in other species. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.

Read More

Answer

No, the Abcam search engine will have recognised the "E" in the search term that you provided. A search for "cathepsin" identifies a number of antibodies to Cathepsin B, D, H & L, but unfortunately, we do not have an antibody to Cathepsin E.

Read More

Question
Answer

This antibody is a polyclonal and will therefore recognise a number of epitopes

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