Revision 8

#96397

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, 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 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.

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).

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 IHC-Bond: IHC Leica Bond 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 G: Goat 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

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 8

CST Logo
Western blot analysis of extracts from various cell lines using FcγRIIB (D8F9C) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Western blot analysis of whole spleen extracts from wild type C57BL/6 mice (lane 1) on Fcgr2b knockout (Taconic Biosciences, Model 580-F) (lane 2) FcγRIIB (D8F9C) XP(r) Rabbit mAb #96397 (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The abscence of signal in the Fcgr2b knockout tissue confirms specificity of the antibody for FcγRIIB.
Western Blotting Image 2: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of FcγRIIB from J774A.1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is FcγRIIB (D8F9C) XP® Rabbit mAb. Western blot analysis was performed using FcγRIIB (D8F9C) XP® Rabbit mAb.
Immunoprecipitation Image 1: FcγRIIB (D8F9C) XP<sup>®</sup> 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 8

CST Logo
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using Fc-gamma-RIIB (D8F9C) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse lung using FcγRIIB (D8F9C) XP® Rabbit mAb.
Immunohistochemistry Image 1: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse forestomach using FcγRIIB (D8F9C) XP® Rabbit mAb.
Immunohistochemistry Image 2: FcγRIIB (D8F9C) XP<sup>®</sup> 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 8

CST Logo
Immunohistochemical analysis of paraffin-embedded mouse placenta using FcγRIIB (D8F9C) XP® Rabbit mAb.
Immunohistochemistry Image 3: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded lung (left) or colon (right) from C57BL/6NTac (wt/wt, model #B6-F, top) and FcγRIIB knockout (-/-, model #580-F, bottom) mice using FcγRIIB (D8F9C) XP® Rabbit mAb. Mice from Taconic Biosciences, Inc.
Immunohistochemistry Image 4: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse spleen using FcγRIIB (D8F9C) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemistry Image 5: FcγRIIB (D8F9C) XP<sup>®</sup> 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 8

CST Logo
Immunohistochemical analysis of paraffin-embedded J774A.1 cell pellet (left, positive) or NIH/3T3 cell pellet (right, negative) using FcγRIIB (D8F9C) XP® Rabbit mAb.
Immunohistochemistry Image 6: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Flow cytometric analysis of mouse splenocytes using FcγRIIB (D8F9C) XP® Rabbit mAb (right) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Samples were co-stained with CD45R (B220)-APC to distinguish the B cell population. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Total viable cells were used for analysis.
Flow Cytometry Image 1: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Flow cytometric analysis of mouse splenocytes using FcγRIIB (D8F9C) XP® Rabbit mAb (right) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Samples were co-stained with CD3-FITC to distinguish the T cell population. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. Total viable cells were used for analysis.
Flow Cytometry Image 2: FcγRIIB (D8F9C) XP<sup>®</sup> 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 8

CST Logo
Flow cytometric analysis of NIH/3T3 cells (blue) and J774A.1 cells (green) using FcγRIIB (D8F9C) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 3: FcγRIIB (D8F9C) XP<sup>®</sup> Rabbit mAb
Flow cytometric analysis of fixed and permeabilized splenocytes from wild type C57BL/6 (top) or Fcgr2b knockout (Taconic Biosciences, Model 580-F) (bottom) mice using FcγRIIB (D8F9C) XP® Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left) co-stained with CD45R/B220 (RA3-6B2) Rat mAb (PE-Cy7® Conjugate) #82922. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
Flow Cytometry Image 4: FcγRIIB (D8F9C) XP<sup>®</sup> 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.