Revision 1

#70168Store 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-Bond, IHC-P, IF-F

REACTIVITY:

M R

SENSITIVITY:

Endogenous

MW (kDa):

30-50

Source/Isotype:

Rabbit IgG

UniProt ID:

#P19437

Entrez-Gene Id:

12482

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
IHC Leica Bond 1:400 - 1:1600
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Frozen) 1:100 - 1:400

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

CD20 (E3N7O) XP® Rabbit mAb recognizes endogenous levels of total CD20 protein.

Species Reactivity:

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of mouse CD20 protein.

Background

B-lymphocyte antigen CD20 (also known as MS4A1; Membrane-spanning 4-domains subfamily A member 1) is a cell surface phosphoprotein involved in the regulation of B cell activation and proliferation (1,2). It is commonly used as a marker to identify B cells and is expressed throughout B cell development, up until their differentiation into plasma cells. CD20 has no known ligand, and its expression and function are largely conserved between human and mouse (1-3). Evidence suggests that CD20 is necessary for store operated calcium (SOC) entry, which leads to elevated cytoplasmic calcium levels required for B cell activation (4,5). Anti-CD20 antibody immunotherapy depletes B cells by activation of the innate monocytic network and is a common treatment for B cell lymphomas, leukemias, and autoimmune diseases (6).

  1. Stashenko, P. et al. (1980) J Immunol 125, 1678-85.
  2. Tedder, T.F. et al. (1985) J Immunol 135, 973-9.
  3. Tedder, T.F. et al. (1988) J Immunol 141, 4388-94.
  4. Bubien, J.K. et al. (1993) J Cell Biol 121, 1121-32.
  5. Li, H. et al. (2003) J Biol Chem 278, 42427-34.
  6. Uchida, J. et al. (2004) J Exp Med 199, 1659-69.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen)

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 1
#70168

CD20 (E3N7O) XP® Rabbit mAb

Western Blotting Image 1: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb(上图)、Myc-Tag (71D10) Rabbit mAb #2278(中图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或转染表达带有 Myc/DDK 标签的全长小鼠 CD20 蛋白(mCD20-Myc/DDK; +)表达载体的 293T 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对不同细胞系和组织的提取物进行蛋白质印迹分析。
Western Blotting Image 3: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对小鼠脾脏、大鼠脾脏和大鼠大脑组织的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: CD20 (E3N7O) XP® Rabbit mAb Expand Image
对小鼠脾组织提取物 CD20 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 CD20 (E3N7O) XP® Rabbit mAb。使用 CD20 (E3N7O) XP® Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunohistochemistry Image 1: CD20 (E3N7O) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx上使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 2: CD20 (E3N7O) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的小鼠肝脏进行免疫组织化学分析。
Immunohistochemistry Image 3: CD20 (E3N7O) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的小鼠脾脏进行免疫组织化学分析。
Immunohistochemistry Image 1: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的 LL/2 同源肿瘤进行免疫组织化学分析。
Immunohistochemistry Image 2: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的 A20 同源肿瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: CD20 (E3N7O) 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 CD20 (E3N7O) XP® Rabbit mAb(左图)对石蜡包埋的小鼠肺脏进行免疫组织化学分析。
Immunohistochemistry Image 4: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb 对石蜡包埋的 A20 细胞沉淀物(左图,阳性)或 LL/2 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb(绿色)和 CD44 (IM7) Rat mAb (Alexa Fluor® 555 Conjugate) #95235(红色)对小鼠肝脏进行共聚焦免疫荧光分析。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
Immunofluorescence Image 2: CD20 (E3N7O) XP® Rabbit mAb Expand Image
使用 CD20 (E3N7O) XP® Rabbit mAb(红色)和 CD3 (17A2) Rat mAb (FITC Conjugate) #86603(绿色)在较低放大倍数(左图)和较高放大倍数(右图)下对小鼠脾脏进行共聚焦免疫荧光分析。在用 Rabbit (DA1E) mAb IgG XP® Isotype Control # 3900 封闭游离二抗结合位点后,随后用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #78060(伪彩青色)标记该组织。