Human B Cell Signaling Flow Cytometry Panel

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:
FC-FP

Reactivity:
H

UniProt ID:
#P15391, #P43405, #P62753, #P08575, #P31749, #P11912

Entrez-Gene Id:
930, 6850, 6194, 5788, 207, 973

Product Usage Information

B cell receptor activation can be induced through treatment with Goat Anti-Human IgM, F(ab')₂ Antibody (Low Endotoxin, Azide-free) #96232. All antibodies in this kit are compatible with the Intracellular Flow Cytometry Kit (Methanol) #13593 and can be used in a single staining mix on fixed and permeabilized cells. Prior to fixation and antibody incubation, we recommend adding a fixable viability dye such as the Ghost Dye Violet 510 Fixable Viability Dye #59863 to enable identification and exclusion of dead cells from analysis.

For individual product dilutions, refer to panel product datasheet or individual product pages.

Gating strategy for observing B cell receptor activation in B cell populations: Apply the same gating strategy for both untreated and treated cell populations. If a fixable viability dye was used, then first gate on viable cells. Next, gate on CD45+ immune cells. Viewing the CD45+ population, gate on singlets using appropriate scatter parameters, such as FSC-A vs. FSC-H. Observe CD19 vs. SSC-A and gate on CD19+ B cells. Observe phospho-CD79A (Tyr182) and phospho-Syk (Tyr525/526) within the CD19+ gate.

Storage

CD45 (HI30) Mouse mAb (violetFluor 450 Conjugate) is supplied in 10 mM NaH₂PO₄, 150 mM NaCl, 0.09% NaN₃, 0.1% gelatin, pH 7.2. CD19 (Intracellular Domain) (D4V4B) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate), Phospho-CD79A (Tyr182) (D1B9) Rabbit mAb (Alexa Fluor^® 488 Conjugate), and Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (PE Conjugate) are supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4^o C. Do not aliquot the antibodies. Protect from light. Do not freeze.

All components in this kit are stable in accordance with the date printed on the outer packaging label when stored at the recommended temperature. Please refer to product labels, datasheets, or web pages for specific “Best By” dates for each individual component.

Specificity/Sensitivity

Each antibody in the Human B Cell Signaling Flow Cytometry Panel detects endogenous levels of its target protein. CD45 (HI30) Mouse mAb (violetFluor 450 Conjugate) detects an epitope within the extracellular domain. CD19 (Intracellular Domain) (D4V4B) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate) detects an epitope within the intracellular domain. Phospho-CD79A (Tyr182) (D1B9) Rabbit mAb (Alexa Fluor^® 488 Conjugate) recognizes endogenous levels of human CD79A protein only when phosphorylated at Tyr188. This corresponds to Tyr182 of mouse CD79A protein. Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (PE Conjugate) detects endogenous levels of Syk protein only when phosphorylated at Tyr525/526 of human Syk. It also detects Syk protein when singly phosphorylated at Tyr526 of human Syk. It does not cross-react with other tyrosine-phosphorylated protein tyrosine kinases.

Source / Purification

Monoclonal antibodies were purified from tissue culture supernatant via affinity chromatography. The purified antibodies were conjugated under optimal conditions, with unreacted dye removed from the preparation.

Description

The Human B Cell Signaling Flow Cytometry Panel can be used to interrogate activation of human B cells among peripheral blood mononuclear cells (PBMCs).

CD45 is a pan leukocyte marker. B cells are identified by expression of CD19. Engagement of the B cell receptor (BCR) triggers the tyrosine phosphorylation of CD79A at Tyr182, which is mediated by Src family kinases. Following engagement of the BCR, Syk is recruited to phosphorylated ITAM motifs within the BCR signaling complex, which facilitates Syk activation through phosphorylation of Tyr525/526 within the kinase activation loop.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

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.

XP is a registered 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.