Overview

  • Product name
    Anti-Apolipoprotein E antibody [E6D7]
    See all Apolipoprotein E primary antibodies
  • Description
    Mouse monoclonal [E6D7] to Apolipoprotein E
  • Host species
    Mouse
  • Specificity
    Clone E6D7 recognizes the E2, E3, and E4 isoforms of Apolipoprotein E.
  • Tested applications
    Suitable for: IHC-FoFr, WB, ELISA, IP, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide spanning the polymorphic amino acid position 158 of ApoE.

  • Positive control
    • IHC-P: Human liver tissue. ICC/IF: HepG2 cells.

Applications

Our Abpromise guarantee covers the use of ab1907 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
IHC-FoFr 1/500.
WB 1/100 - 1/1000. Predicted molecular weight: 38 kDa.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 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

  • All lanes : Anti-Apolipoprotein E antibody [E6D7] (ab1907) at 1/1000 dilution

    Lane 1 : Human THP-1 macrophage whole cell lysate - treated with 100 µg/ml oxidized LDL (oxLDL)
    Lane 2 : Human THP-1 macrophage whole cell lysate - treated with 100 µg/ml oxidized LDL (oxLDL) + 10 µg/ml apoA-I

    Lysates/proteins at 70 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 38 kDa


    Exposure time: 4 minutes


    10% SDS -PAGE.

    Blocked with 5% milk for 2 hours at 22°C.

    Incubated with the primary antibody for 18 hours at 4°C in TBST + 5% milk.

    See Abreview

  • Ab1907 staining Human liver. Staining is localised to cytoplasmic and extracelular regions.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • IHC-FoFr image of Apolipoprotein E staining on choroid plexus of APP transgenic mouse brain sections using ab1907 (1:500). The mouse was perfused with 4% PFA in 0.1M PBS. The sections were permeabilised using 0.1% TriktonXin 0.1% PBS. 10% serum was used for 1 hour at 24°C for blocking step. The sections were stained using ab1907 at 1:500 dilution and Rabbit monoclonal to anti-mouse IgG conjugated to Alexa Fluor® 568 was used at 1:1000 to detect the primary antibody. DAPI was used to stain the nuclei.

    See Abreview

  • ICC/IF image of ab1907 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat 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 (ab1907, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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:
  • Main BS  et al. Apolipoprotein E4 impairs spontaneous blood brain barrier repair following traumatic brain injury. Mol Neurodegener 13:17 (2018). Read more (PubMed: 29618365) »
  • Hondius DC  et al. Proteomics analysis identifies new markers associated with capillary cerebral amyloid angiopathy in Alzheimer's disease. Acta Neuropathol Commun 6:46 (2018). Read more (PubMed: 29860944) »
See all 7 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Yong Sun Lee

Verified customer

Submitted Jul 03 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (1 lane : A549 / 2 lane : H460 human lung cancer c)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
1 lane : A549 / 2 lane : H460 human lung cancer c
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Mr. Yong Sun Lee

Verified customer

Submitted May 22 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Human THP-1 macrophages)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
70 µg
Treatment
100 µg/ml oxidized LDL (oxLDL) +/- 10 µg/ml apoA-I
Specification
Human THP-1 macrophages
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Anca Sima

Verified customer

Submitted Jul 03 2015

Question
Answer

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

I am sorry to confirm that as far as we are aware, ab1907 Apolipoprotein E antibody [E6D7] has not yet been tested with samples from rat. All tested species covered by the 6 month guarantee are stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

I have checked the alignment of the immunogen with the rat protein, this is around 75%. We would usually suggest that alignment shoudl be over 85% to predict that an antibody will detect in a different species. The alignment with the rat is still reasonably high in this case and there may be some crossreactivity. However, we are not able to guarantee this without further testing.

If you would like to test ab1907in rat, please do not hesitaet to contact me again by replying to this message prior to the purchase as you would be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

Read More

Answer

Merci de nous avoir contactés.

Je suis vraiment désolée pour le délai de ma réponse, je suis toujours en contact avec notre collaborateur a propos de votre question.

Cependant, Je peux vous confirmer que l’acide aminé en position 158 est un histidine. Cet anticorps peut reconnaitre les isoforms E2, E3 et E4 de la protéine apolipoprotein E.

Une fois de plus, je suis désolée pour cette réponse tardive et je vous tiendrai au courant si j’ai d’autres informations à propos de ce sujet.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Monoclonal de lapin gratuit pour tout achat d'anticorps primaire, dans la limite des stocks disponibles. Notez le code "RABMAB-XBSMG" sur votre prochaine commande d’anticorps primaire. Pour plus d'informations, visitez le lien suivant: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Question
Answer

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 for one vial of ab20874.

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
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain lysate)
Loading amount
20 µg
Specification
Brain lysate
Gel Running Conditions
Reduced Denaturing (12)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Aug 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Cell (Brain section)
Specification
Brain section
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Aug 07 2012

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 (Thyroid)
Specification
Thyroid
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1%

Abcam user community

Verified customer

Submitted Jan 29 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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