Overview

  • Product name
    Anti-Apolipoprotein E antibody [D6E10]
    See all Apolipoprotein E primary antibodies
  • Description
    Mouse monoclonal [D6E10] to Apolipoprotein E
  • Host species
    Mouse
  • Specificity
    Mouse reactivity: Please be aware that we have received positive as well as negative feedback for reactivity of this antibody with mouse samples. The antibody is not being batch-tested in the mouse samples. Anti-Apolipoprotein E antibody [D6E10] recognizes the E2, E3 and E4 isoforms of apolipoprotein E. It was raised against a peptide sequence corresponding to aa 141-160 of human Apo-E.
  • Tested applications
    Suitable for: ICC/IF, IP, WB, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Apolipoprotein E aa 141-160.
    Sequence:

    QAMLGQSTEE LRVRLASHLR

  • General notes

    This product was changed from ascites to tissue culture supernatant on 2nd February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Applications

Our Abpromise guarantee covers the use of ab1906 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. PubMed: 21163940
IP Use at an assay dependent concentration. PubMed: 21163940
WB 1/1000 - 1/10000. Predicted molecular weight: 38 kDa.
IHC-P 1/1000 - 1/100000. Antigen retrieval is not essential but may optimise staining.

The staining intensity of formalin-fixed paraffin embedded tissues may be significantly improved by pretreatment methods such as: 70% Formic acid for 10-30 minutes at room temperature or Hydrolytic autoclaving.

IHC-Fr 1/1000 - 1/100000.
Flow Cyt Use at an assay dependent concentration. PubMed: 22896615

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

  • All lanes : Anti-Apolipoprotein E antibody [D6E10] (ab1906) at 1/500 dilution

    All lanes : SCC61 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-mouse IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 60 kDa (possible non-specific binding)


    Exposure time: 2 minutes


    Blocked with 5% milk for 1 hour at 22°C.

    Incubated with the primary antibody in 5% milk for 16 hours at 4°C.

    See Abreview

  • ab1906 staining Apolipoprotein E in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/200 in TBS/BSA/azide) for 2 hours at 21°C. An undiluted biotin-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab1906 staining Apolipoprotein E in zebrafish retina sections by IHC-Fr. The tissue was fixed with paraformaldehyde and an antigen retrieval step was performed with sodium citrate pH 6. Blocking of the sample was done with 5% BSA in PBS containing 01% Tween 20 and 0.5% Triton X, for 60 minutes at 23°C, followed by staining with ab1906 at 1/100 in blocking solution for 16h at 4°C. An Alexa Fluor® 647 conjugated goat anti-mouset polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei are stained in blue with DAPI. Apolipoprotein E expression can be observed in Muller cells.

    See Abreview

References

This product has been referenced in:
  • Quartey MO  et al. Alzheimer Disease and Selected Risk Factors Disrupt a Co-regulation of Monoamine Oxidase-A/B in the Hippocampus, but Not in the Cortex. Front Neurosci 12:419 (2018). Read more (PubMed: 29997470) »
  • Llobet L  et al. Pharmacologic concentrations of linezolid modify oxidative phosphorylation function and adipocyte secretome. Redox Biol 13:244-254 (2017). WB . Read more (PubMed: 28600981) »
See all 28 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (SCC61 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
SCC61 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 20 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (Muller cell, retina)
Specification
Muller cell, retina
Permeabilization
Yes - 0.1% Triton X
Fixative
Paraformaldehyde

Dr. Ryan Macdonald

Verified customer

Submitted Nov 08 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Serum (serum)
Loading amount
100 µg
Specification
serum
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Feb 15 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 getting back to me.

This is to let you know that I have placed a new order for you - for one vial of ab1906 as a free of charge replacement and the new order number is 1206066.

I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product.

Read More

Answer

Thank you for getting back to us with the updates.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Currently we have new batch in our system so I can offer your troubled customer a free of charge replacement and a credit note which can be used in the next order.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Question
Answer

Thank you for contacting us and for sending the questionnaire. 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. The only lot of ab17929 we have in stock is the same as the one metioned in the questionaaire. Therefore, as requested, I have issued a free of charge replacement with one unit of ab20874. This free of charge replacement was added to the order number ******.

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

I looked at the protocol and I would like to suggest:
- to load a maximum of 30µg on the gel. Too much protein usually brings a lot of background.
- to dilute the primary antibody more (1/1000 is commonly used), too much antibody usually brings a lot of background too.

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.

Read More

Question

Hello,

I’ve got the fullfilled document from my client and I send it back to you. I hope that it helps to solve the problem.

AB1906

2) Abcam order reference number or product batch number



3) Description of the problem

Antibody binds unspecific



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…)

Tissue (liver) proteins

Lysis buffer

20mM Hk_HEPES = 2mM CaCl2

Protease inhibitors:

Protease mux (GE Healthcare)

Phosphatase inhibitors

Reducing agent

SDS

Boiling for ≥5 min? yes/no

Protein loaded ug/lane or cells/lane 10ug

Positive control

Negative control



5) Percentage of gel 12%

Type of membrane PVDF

Protein transfer verified marker of size

Blocking agent and concentration 1% BSA

Blocking time overnight

Blocking temperature 4°C



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Anti-Apolipoprotein E antibody

Concentration or dilution 1/500

Diluent buffer 1% BSA in PBS/Tween

Incubation time 1,5h

Incubation temperature: RT



7) Secondary antibody: http://www.sigmaaldrich.com/catalog/product/sigma/a9044?lang=pl&region=PL

Species:

Reacts against: mouse

Concentration or dilution 1/80000

Diluent buffer 1% BSA in PBS/Tween

Incubation time 1h

Incubation temperature: RT

Fluorochrome or enzyme conjugate: HRP



8) Washing after primary and secondary antibodies:

Buffer PBS tween

Number of washes 4 washes by7min



9) Detection method



10) How many times have you run this staining?

twice

Do you obtain the same results every time?

yes

What steps have you altered to try and optimize the use of this antibody?

Used more antibody –dilution 1/300!



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.



I attached both blots I get for this antibody



1/500







1/300



Best regards

Read More
Answer

Thank you for your enquiry regardingab1906 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer ishaving problems with this antibody.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

1) Blocking:

I would suggest increasing the concentration of the blocking agent from 1% to 5% and using this solution for diluting the primary and the secondary antibody as well.

2) Dilution of the primary antibody:

It may be worth diluting the primary antibody further ie. 1/750 and 1/1000.

3) No primary control:

Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not? If you have not done yet, I would advise you to check it.

I hope this will be useful for you. Should your customerstill have any problem with this antibody after following these suggestions, then please confirm the batch number and the Abcam Order Number and do not hesitate to contact our Technical Department again.

Read More

Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.

1) Could you provide some further details of the protocol used and complete the following form (attached as a word document). I am particularly interested in the following information:

- sample preparation,

- lysis buffer used,

- specification of the secondary antibody (host species, specificity),

- positive control applied

2) It would be much appreciated if you could attach a full size WBimage with molecular marker bands (MW) to the response. Please also describe what the different lanes represent.

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

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Testis)
Specification
Testis
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Sep 07 2012

1-10 of 20 Abreviews or Q&A

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