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FastScan Total Btk ELISA Kit #36609

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ELISA Image 1: FastScan™ Total Btk ELISA Kit
Figure 1. Btk protein is expressed in Ramos cells but absent in HeLa cells. The relationship between lysate protein concentration from Ramos and HeLa cells and the absorbance at 450 nm using the FastScan™ Total Btk ELISA Kit #36609 is shown in the upper figure. The corresponding western blot using a Btk antibody is shown in the lower figure. Unstarved Ramos (1.0 x106 cells/ml) or HeLa (90% confluence) cells were harvested and then lysed.

To Purchase # 36609**

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • Product Includes
  • Related Products
Product IncludesVolume (with Count)Solution Color
FastScan™ ELISA Microwell Strip Plate, 96 Well #532571 x 96 tests
Btk Mouse Capture mAb #765451 x 1 eaGreen (Lyophilized)
Btk Rabbit HRP-linked mAb #731011 x 1 eaRed (Lyophilized)
FastScan™ ELISA Capture Antibody Diluent #160761 x 3 mlGreen
FastScan™ ELISA HRP Antibody Diluent #281201 x 3 ml
TMB Substrate #70041 x 11 ml
STOP Solution #70021 x 11 ml
Sealing Tape #545031 x 1 ea
ELISA Wash Buffer (20X) #98011 x 25 ml
FastScan™ ELISA Cell Extraction Buffer (5X) #699051 x 10 ml
FastScan™ ELISA Cell Extraction Enhancer Solution (50X) #252431 x 1 ml
FastScan™ ELISA Kit #36609 Positive Control Type 2 #646031 x 1 ea

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.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

The FastScan™ Total Btk ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Btk. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with Btk in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of Btk.

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

Protocol

Specificity / Sensitivity

The FastScan™ Total Btk ELISA Kit detects endogenous levels of Btk as 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.

Species Reactivity:

Human

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).

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.
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