Revision 5

#29765Store 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-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

100-140

Source/Isotype:

Rabbit IgG

UniProt ID:

#P05067

Entrez-Gene Id:

351

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:12800 - 1:25600

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

APP (E8B3O) XP® Rabbit mAb recognizes endogenous levels of total APP protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro620 of human APP protein.

Background

Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (1). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (2-5). Phosphorylation at Thr668 (a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell-cycle dependent and peaks during G2/M phase (4). APP phosphorylated at Thr668 exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).

  1. Selkoe, D.J. (1996) J Biol Chem 271, 18295-8.
  2. Caporaso, G.L. et al. (1992) Proc Natl Acad Sci USA 89, 3055-9.
  3. Hung, A.Y. and Selkoe, D.J. (1994) EMBO J 13, 534-42.
  4. Suzuki, T. et al. (1994) EMBO J 13, 1114-22.
  5. Ando, K. et al. (1999) J Neurosci 19, 4421-7.
  6. Iijima, K. et al. (2000) J Neurochem 75, 1085-91.

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 IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) 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.
SignalStain is a registered 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.

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
#29765

APP (E8B3O) XP® Rabbit mAb

Western Blotting Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® Rabbit mAb(上图)与 β-Actin (D6A8) Rabbit mAb #8457(下图),对 A549 shControl 和 A549 shAPP 细胞提取物进行蛋白印迹分析。
Western Blotting Image 2: APP (E8B3O) XP® Rabbit mAb Expand Image
Western blot analysis of HAP1 extracts from WT (left) or APP KO (right) using APP (E8B3O) XP® Rabbit mAb. 用 Ponceau S 染色的膜进行总蛋白标准化(下图)。这些数据由 YCharOS Inc. 提供,该公司是一家开放科学公司,作为 CST 科学家生成的验证数据的合作伙伴,其使命是表征市售抗体。
Immunoprecipitation Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
对 A549 shGFP 细胞提取物的 APP 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 APP (E8B3O) XP® Rabbit mAb。使用 APP (E8B3O) XP® Rabbit mAb 进行蛋白质印迹法分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunohistochemistry Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® 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 2: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® Rabbit mAb 对石蜡包埋的正常人脑组织进行免疫组织化学分析。
Immunohistochemistry Image 3: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® Rabbit mAb 对已经过 shRNA 处理的对照品(左图)或已经过 shRNA 处理的 APP(右图)的石蜡包埋的 A549 细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 4: APP (E8B3O) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 APP (E8B3O) XP® Rabbit mAb(左图)对石蜡包埋的人卵巢浆液性癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 5: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® Rabbit mAb(上图)或 APP (E3F3P) Rabbit mAb #19389(下图) 对石蜡包埋的人脾细胞(左图)或气管腺样囊性癌细胞(右图)进行免疫组织化学分析。这两种抗体可检测人 APP上的独立、独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 6: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E8B3O) XP® Rabbit mAb 对石蜡包埋的人前列腺癌进行免疫组织化学分析。
Immunofluorescence Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
使用 APP (E9M5J) Rabbit mAb(绿色)和 Rab5 (E6N8S) Mouse mAb #46449 (红色)对表达非靶向 shRNA(左,阳性)或 shAPP(右,阴性)的 A549 细胞进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。