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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 5
#39327

ApoE4 (E5M4L) Rabbit mAb

Western Blotting Image 1: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb(上图)和 ApoE (pan) (D7I9N) Rabbit mAb #13366(下图)对重组 ApoE2、ApoE4 和 ApoE3 蛋白 (5ng) 进行蛋白印迹分析。
Western Blotting Image 2: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb(上图)、ApoE (pan) (D7I9N) Rabbit mAb #13366(中间)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或转染表达全长人 ApoE4 蛋白 (hApoE4; +)、全长人 ApoE3 蛋白 (hApoE3; +) 或全长人 ApoE2 蛋白 (hApoE2; +) 的表达载体的 293T 细胞的提取物进行蛋白印迹分析。
Western Blotting Image 3: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对 HepG2 细胞提取物(泳道 1)、转染空载(泳道 2)或瞬时转染 ApoE4(泳道 3)的 293T 细胞提取物、野生型 C57BL/6NTac 模型 #B6-F 小鼠(泳道 4)或 ApoE4 敲入型小鼠(模型 1549-F)(泳道 5)的全肝提取物进行蛋白质印迹分析。Taconic Biosciences, Inc. 的小鼠
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. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 2-40 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。

Immunohistochemistry Image 1: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对石蜡包埋的人肝细胞癌进行免疫组织化学分析。
Immunohistochemistry Image 2: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对石蜡包埋的人尿路上皮癌进行免疫组织化学分析。
Immunohistochemistry Image 3: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对来自 C57BL/6NTac(wt/wt,model #B6-F,上图)或 APOE4 敲入型(hu/hu,模型#1549-F,下图)且石蜡包埋的肝脏(左图或脑(右图)进行免疫组织化学分析。Taconic Biosciences, Inc. 的小鼠
Immunohistochemistry Image 4: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对石蜡包埋的正常人肺进行免疫组织化学分析。
Immunohistochemistry Image 5: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对石蜡包埋的人成熟卵巢畸胎瘤进行免疫组织化学分析。
Immunohistochemistry Image 6: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb 对石蜡包埋的人肺腺癌进行免疫组织化学分析。
Immunohistochemistry Image 7: ApoE4 (E5M4L) Rabbit mAb Expand Image
使用 ApoE4 (E5M4L) Rabbit mAb(顶部小图)或 ApoE (pan) (D7I9N) Rabbit mAb #13366(底部小图)对对照或转染人 ApoE 同工型 ApoE4、ApoE2 和 ApoE3(从左到右,如图所示)的石蜡包埋的 293T 细胞沉淀物进行免疫组织化学分析。注意,与过表达的 ApoE2 和 ApoE3 的交叉反应性有限。在内源性模型中,不太可能与这些同工型有交叉反应性。