PathScan^® Phospho-Btk (Tyr223) Sandwich ELISA kit

1 Kit

(96 assays)

Species Cross Reactivity:
H

UniProt ID:
#Q06187

Entrez-Gene Id:
#695

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.

Product Includes	Product #	Quantity	Color	Storage Temp
Btk Mouse mAb Coated Microwells	60976	96 tests		+4C
P-Btk (Tyr223) Rabbit Detection mAb	88236	1 ea	Green (Lyophilized)	+4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)	13272	1 ea	Red (Lyophilized)	+4C
Detection Antibody Diluent	13339	11 ml	Green	+4C
HRP Diluent	13515	11 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
ELISA Sample Diluent	11083	25 ml	Blue	+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The PathScan^® Phospho-Btk (Tyr223) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Btk protein phosphorylated at Tyr223. A Btk mouse antibody has been coated onto the microwells. After incubation with cell lysates, both phospho- and non-phospho-Btk proteins are captured by the coated antibody. Following extensive washing, a phospho-Btk (Tyr223) rabbit antibody is added to detect the captured phospho-Btk protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Btk phosphorylated at Tyr223.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan^® Phospho-Btk (Tyr223) Sandwich ELISA kit detects endogenous levels of Btk protein phosphorylated at Tyr223 in human cells. This kit sensitivity is shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Bruton's tyrosine kinase (Btk) is a member of the Btk/Tec family of cytoplasmic tyrosine kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain and Src homology SH3 and SH2 domains. Btk plays an important role in B cell development (1,2). Activation of B cells by various ligands is accompanied by Btk membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate (3-5). The membrane-localized Btk is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinases, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full activation (6,7). The activation of Btk is negatively regulated by PKCβ through phosphorylation of Btk at Ser180, which results in reduced membrane recruitment, transphosphorylation, and subsequent activation (8). The PKC inhibitory signal is likely to be a key determinant of the B cell receptor signaling threshold to maintain optimal Btk activity (8).

Background References

Trademarks and Patents

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

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

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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