Revision 6

#2756

Store at -20C

Numb (C29G11) Rabbit mAb

Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

[email protected]

cellsignal.com

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, IF-F, IF-IC

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
72, 74

Source/Isotype:
Rabbit IgG

UniProt ID:
#P49757

Entrez-Gene Id:
8650

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:100
Immunofluorescence (Frozen)	1:800 - 1:1600
Immunofluorescence (Immunocytochemistry)	1:1600 - 1:3200

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

Numb (C29G11) Rabbit mAb detects endogenous levels of total Numb protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala570 of human Numb protein.

Background

Numb contains an amino-terminal phosphotyrosine binding (PTB) domain and carboxy-terminal endocytic binding motifs for α-adaptin and EH (Eps15 homology) domain-containing proteins, indicating a role in endocytosis (1,2). There are four mammalian Numb splicing isoforms that are differentially expressed and may have distinct functions (3-5). Numb acts as a negative regulator of Notch signaling by promoting ubiquitination and degradation of Notch (6). The protein is asymmetrically segregated into one daughter cell during cell division, producing two daughter cells with different responses to Notch signaling and different cell fates (7,8). The localization of Numb can also be regulated by G protein-coupled receptor (GPCR) and protein kinase C (PKC) signaling (9).

Background References

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

W: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com

For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#2756

Numb (C29G11) Rabbit mAb

Western blot analysis of extracts from various cell lines using Numb (C29G11) Rabbit mAb.

Western Blotting Image 1: Numb (C29G11) Rabbit mAb

Immunoprecipitation/western blot analysis of lysates from HeLa cells. Lane 1 contains lysate input (10%), lane 2 was immunoprecipitated with non-specific rabbit IgG (#3900), lane 3 was immunoprecipitated with Numb (C29G11) Rabbit mAb #2756. Western blot analysis was performed using Numb (C29G11) Rabbit mAb #2756.

Immunoprecipitation Image 1: Numb (C29G11) Rabbit mAb

Confocal immunofluorescent analysis of fixed frozen mouse testis using Numb (C29G11) Rabbit mAb (green) and ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 1: Numb (C29G11) Rabbit mAb

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com

For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#2756

Numb (C29G11) Rabbit mAb

Confocal immunofluorescent analysis of fixed frozen mouse brain labeled with Numb (C29G11) Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red), and ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 2: Numb (C29G11) Rabbit mAb

Confocal immunofluorescent analysis of fixed frozen mouse skeletal muscle using Numb (C29G11) Rabbit mAb (green) and ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 3: Numb (C29G11) Rabbit mAb

Confocal immunofluorescent analysis of fixed frozen mouse kidney using Numb (C29G11) Rabbit mAb (green) and ProLong^® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 4: Numb (C29G11) Rabbit mAb

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com

For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#2756

Numb (C29G11) Rabbit mAb

Confocal immunofluorescent analysis of HeLa cells using Numb (C29G11) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Confocal immunofluorescent analysis of control HCT 116 cells (left) or CRISPR/Cas9 NUMB knockdown HCT 116 cells (right) using Numb (C29G11) Rabbit mAb (green) and DAPI #4083 (blue).

Confocal immunofluorescent analysis of HeLa cells, mock transfected (left, high-expressing) or transfected with siRNA directed against NUMB (right, low-expressing), using Numb (C29G11) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com

For Research Use Only. Not for Use in Diagnostic Procedures.