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.

使用に関する制限

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研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

Revision 5
#29765

APP (E8B3O) XP® Rabbit mAb

Western Blotting Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from A549 shControl and A549 shAPP cells using APP (E8B3O) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
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. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
Immunoprecipitation Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunoprecipitation of APP from A549 shGFP cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is APP (E8B3O) XP® Rabbit mAb. Western blot analysis was performed usingAPP (E8B3O) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Immunohistochemistry Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using APP (E8B3O) XP® Rabbit mAb.
Immunohistochemistry Image 2: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human brain using APP (E8B3O) XP® Rabbit mAb.
Immunohistochemistry Image 3: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded A549 cell pellet, control shRNA-treated (left) or APP shRNA-treated (right), using APP (E8B3O) XP® Rabbit mAb.
Immunohistochemistry Image 4: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using APP (E8B3O) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 5: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human spleen (left) or adenoid cystic carcinoma of the trachea (right) using APP (E8B3O) XP® Rabbit mAb (top) or APP (E3F3P) Rabbit mAb #19389 (bottom). These two antibodies detect independent, unique epitopes on human APP. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 6: APP (E8B3O) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using APP (E8B3O) XP® Rabbit mAb.
Immunofluorescence Image 1: APP (E8B3O) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of A549 cells expressing either a non-targeting shRNA (left, positive) or shAPP (right, negative) using APP (E9M5J) Rabbit mAb (green) and Rab5 (E6N8S) Mouse mAb #46449 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).