Revision 6

#13366Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-F, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

35

Source/Isotype:

Rabbit IgG

UniProt ID:

#P02649

Entrez-Gene Id:

348

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:250 - 1:1000
Immunofluorescence (Frozen) 1:800 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:3200 - 1:6400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #10197.

Specificity / Sensitivity

ApoE (pan) (D7I9N) Rabbit mAb recognizes endogenous levels of total ApoE protein. This antibody also recognizes overexpressed ApoE2, ApoE3 and ApoE4 proteins.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro285 of human ApoE protein.

Background

Apolipoproteins are plasma lipoproteins that function as transporters of lipids and cholesterol in the circulatory system. Chylomicrons are a fundamental class of apolipoproteins containing very low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL) (1,2).

Human ApoE has three isoforms: ApoE2, ApoE3, and ApoE4. These three isoforms differ in the combination of cysteine and arginine residues located at positions 130 and 176. The ApoE4 isoform contains arginine at both locations. Research studies have linked ApoE4 function to neuronal plasticity, synaptogenesis, and neurodegenerative diseases (3). ApoE4 is produced in the liver and brain, although it is widely expressed in other tissues, such as the lung, spleen, and ovary. Investigators have established the ApoE4 allele as a genetic risk factor for Alzheimer’s disease (AD), accounting for 50-60% of the genetic variation in the disease (4). Research studies indicate that patients expressing ApoE4 have a reduced capacity for synaptic plasticity, an earlier age of onset of AD, and an increase in amyloid-beta (Aβ) deposition. The increase in Aβ suggests a role for ApoE4 in the impairment of amyloid clearance (5).

  1. Kwiterovich, P.O. (2000) Am J Cardiol 86, 5L-10L.
  2. Hussain, M.M. (2000) Atherosclerosis 148, 1-15.
  3. Raber, J. et al. Neurobiol Aging 25, 641-50.
  4. Corder, E.H. et al. (1993) Science 261, 921-3.
  5. Holtzman, D.M. et al. (2000) Proc Natl Acad Sci U S A 97, 2892-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 6
#13366

ApoE (pan) (D7I9N) Rabbit mAb

Western Blotting Image 1: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Western blot analysis of cell extracts from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min) and human cerebellum using ApoE (pan) (D7I9N) Rabbit mAb.
Western Blotting Image 2: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Western blot analysis of cell extracts from 293T cells, treated with Brefeldin A #9972 (10 ng/ml, 90 min; +) and mock transfected (-) or transfected with a construct expressing full-length human ApoE2 (hApoE2; +), ApoE3 (hApoE3; +), or ApoE4 (hApoE4; +), using ApoE (pan) (D7I9N) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western Blotting Image 3: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Western blot analysis of extracts from HepG2 cells (lane 1), 293T mock transfected (lane 2) or transiently transfected with a construct expressing ApoE4 (lane 3), whole liver extracts from wild type C57BL/6NTac model #B6-F mice (lane 4), or ApoE4 knock-in (model #1549-F) (lane 5), whole brain extracts from wild type C57BL/6NTac mice (lane 6), or ApoE4 knock-in (lane 7) using ApoE (pan) (D7I9N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Mice from Taconic Biosciences, Inc.
Immunoprecipitation Image 1: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunoprecipitation of ApoE from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ApoE (pan) (D7I9N) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemistry Image 1: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human kidney using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemistry Image 2: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded liver (left) or brain (right) from C57BL/6NTac (wt/wt, model #B6-F, top) and APOE4 knock-in (hu/hu, model #1549-F, bottom) mice using ApoE (pan) (D7I9N) Rabbit mAb. Mice from Taconic Biosciences, Inc.
Immunohistochemistry Image 3: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human spleen using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemistry Image 4: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human small intestine using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemistry Image 5: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human skin using ApoE (pan) (D7I9N) Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).
Immunohistochemistry Image 6: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left-top) or transfected with human ApoE isoforms ApoE2 (right-top), ApoE3 (left-bottom), and ApoE4 (right-bottom) using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemistry Image 7: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Hep G2 cell pellet (left, positive) or 293T cell pellet (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb.
Immunofluorescence Image 1: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen hippocampus from C57BL/6 (left) and APOE4 knock-in (hu/hu, model #1549-F, right) mice labeled with ApoE (pan) (D7I9N) Rabbit mAb (top, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with GFAP (GA5) Mouse mAb (Alexa Fluor® 594 Conjugate) #8152 (bottom, red), and AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #89851 (bottom, blue pseudocolor). Mice from Taconic Biosciences, Inc.
Immunofluorescence Image 1: ApoE (pan) (D7I9N) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of Hep G2 cells (left, positive) and 293T cells (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb #13366 (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).