Revision 5

#39327Store 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, W-S, IHC-P

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
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:400 - 1:1600

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.

Specificity / Sensitivity

ApoE4 (E5M4L) Rabbit mAb recognizes endogenous levels of total ApoE4 protein. This antibody does not cross-react with ApoE2 or ApoE3 by western blot and is not expected to cross-react with endogenous ApoE2 or ApoE3 by immunohistochemistry. Non-specific staining was observed in kidney by immunohistochemistry.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg130 of human ApoE4 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. (2004) 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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IHC-P: Immunohistochemistry (Paraffin)

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.
SignalStain 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 5
#39327

ApoE4 (E5M4L) Rabbit mAb

Western Blotting Image 1: ApoE4 (E5M4L) Rabbit mAb Expand Image
Western blot analysis of recombinant ApoE2, ApoE4, and ApoE3 proteins (5ng) using ApoE4 (E5M4L) Rabbit mAb (upper) and ApoE (pan) (D7I9N) Rabbit mAb #13366 (lower).
Western Blotting Image 2: ApoE4 (E5M4L) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human ApoE4 protein (hApoE4; +), full-length human ApoE3 protein (hApoE3; +), or full-length human ApoE2 protein (hApoE2; +), using ApoE4 (E5M4L) Rabbit mAb (upper), ApoE (pan) (D7I9N) Rabbit mAb #13366 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 3: ApoE4 (E5M4L) 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) using ApoE4 (E5M4L) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Mice from Taconic Biosciences, Inc.
Western Blotting Image 1: ApoE4 (E5M4L) Rabbit mAb Expand Image

Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells transfected with constructs expressing full-length human ApoE4 protein using ApoE4 (E5M4L) Rabbit mAb #39327. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 2-40 kDa separation module.

Immunohistochemistry Image 1: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemistry Image 2: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemistry Image 3: ApoE4 (E5M4L) 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 ApoE4 (E5M4L) Rabbit mAb. Mice from Taconic Biosciences, Inc.
Immunohistochemistry Image 4: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human lung using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemistry Image 5: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human mature ovarian teratoma using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemistry Image 6: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemistry Image 7: ApoE4 (E5M4L) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control or transfected with human ApoE isoforms ApoE4, ApoE2, and ApoE3 (from left to right as indicated), using ApoE4 (E5M4L) Rabbit mAb (top panels) or ApoE (pan) (D7I9N) Rabbit mAb #13366 (bottom panels). Note the limited cross-reactivity with overexpressed ApoE2 and ApoE3. Cross-reactivity with these isoforms in endogenous models is unlikely.