Revision 8

#96397Store 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, FC-FP

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

60-80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08101

Entrez-Gene Id:

14130

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:100
Immunohistochemistry (Paraffin) 1:100
Flow Cytometry (Fixed/Permeabilized) 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.

For a carrier free (BSA and azide free) version of this product see product #81563.

Specificity / Sensitivity

FcγRIIB (D8F9C) XP® Rabbit mAb recognizes endogenous levels of total FcγRIIB protein. Please note that non-specific signal was observed in kidney tubules by IHC.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro284 of mouse FcγRIIB protein.

Background

FcγRIIB (CD32B) is a low affinity, IgG Fc-binding receptor expressed on B cells, monocytes, macrophages, and dendritic cells (DCs) (1-3). It is the inhibitory Fc receptor and signals through an immunoreceptor tyrosine-based inhibitory motif (ITIM) within its carboxy-terminal cytoplasmic tail (2). Binding of immune complexes to FcγRIIB results in tyrosine phosphorylation of the ITIM motif at Tyr292 and recruitment of the phosphatase SHIP, which mediates inhibitory effects on immune cell activation (2,4). In this way, FcγRIIB suppresses the effects of activating Fc-binding receptors (3). For example, mice deficient for FcγRIIB have greater T cell and DC responses following injection of immune complexes (5,6). In addition, FcγRIIB plays a role in B cell affinity maturation (7). Signaling through FcγRIIB in the absence of signaling through the B cell receptor (BCR) is proapoptotic, while signaling through FcγRIIB and the BCR simultaneously attenuates the apoptotic signal and results in selection of B cells with higher antigen affinity (7).

  1. Tridandapani, S. et al. (2002) J. Biol. Chem. 277, 5082-89.
  2. Tridandapani, S. et al. (1997) Mol. Cell. Biol. 17, 4305-11.
  3. Guilliams, M. et al. (2014) Nat Rev Immunol 14, 94-108.
  4. Bruhns, P. et al. (2000) J. Biol. Chem. 275, 37357-64.
  5. Kalergis, A.M. and Ravetch, J.V. (2002) J Exp Med 195, 1653-9.
  6. Desai, D.D. et al. (2007) J Immunol 178, 6217-26.
  7. Pearse, R.N. et al. (1999) Immunity 10, 753-60.

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-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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 8
#96397

FcγRIIB (D8F9C) XP® Rabbit mAb

Western Blotting Image 1: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对来自各种细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 2: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP(r) Rabbit mAb #96397(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图),对来自野生型 C57BL/6 小鼠(泳道 1)或 Fcgr2b 敲除小鼠(Taconic Biosciences,型号 580-F)(泳道 2)的完整脾脏提取物进行蛋白质印迹分析。Fcgr2b 敲除组织中信号的缺失证实了该抗体对 FcγRIIB 的特异性。
Immunoprecipitation Image 1: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
对 J774A.1 细胞提取物 FcγRIIB 进行免疫沉淀法分析。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 FcγRIIB (D8F9C) XP® Rabbit mAb。使用 FcγRIIB (D8F9C) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Fc-gamma-RIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的 LL/2 同基因肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 1: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的小鼠肺组织进行免疫组织化学分析。
Immunohistochemistry Image 2: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的小鼠前胃组织进行免疫组织化学分析。
Immunohistochemistry Image 3: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的小鼠胎盘组织进行免疫组织化学分析。
Immunohistochemistry Image 4: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的来自 C57BL/6NTac 小鼠(wt/wt,型号#B6-F,上图)和 FcγRIIB 敲除小鼠(-/-,型号 #580-F,下图)的肺组织(左图)或结肠组织(右图)进行免疫组织化学分析。Taconic Biosciences, Inc. 的小鼠
Immunohistochemistry Image 5: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
在对照肽(左图)或抗原特异性肽(右图)存在的情况下,使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的小鼠脾组织进行免疫组织化学分析。
Immunohistochemistry Image 6: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb 对石蜡包埋的 J774A.1 细胞沉淀物(左图,阳性)或 NIH/3T3 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Flow Cytometry Image 1: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)比较,使用 FcγRIIB (D8F9C) XP® Rabbit mAb(右图)对小鼠脾细胞进行流式细胞分析。样品使用 CD45R (B220)-APC 共染色,以区分 B 细胞群。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。所有活细胞均用于分析。
Flow Cytometry Image 2: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)比较,使用 FcγRIIB (D8F9C) XP® Rabbit mAb(右图)对小鼠脾细胞进行流式细胞分析。样品使用 CD3-FITC 共染色,以区分 T 细胞群。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 作为二抗。所有活细胞均用于分析。
Flow Cytometry Image 3: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 NIH/3T3 细胞(蓝色)和 J774A.1 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Flow Cytometry Image 4: FcγRIIB (D8F9C) XP® Rabbit mAb Expand Image
使用 FcγRIIB (D8F9C) XP® Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染色的 CD45R/B220 (RA3-6B2) Rat mAb (PE-Cy7® Conjugate) #82922, 对固定和透化的来自野生型 C57BL/6 小鼠(上图)或 Fcgr2b 敲除小鼠(Taconic Biosciences,模型 580-F )(下图)的脾细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 作为二抗。