PE Mouse Anti-Btk (pY551)/Itk (pY511)

BD Phosflow™ PE Mouse Anti-Btk (pY551)/Itk (pY511)

Name: PE Mouse Anti-Btk (pY551)/Itk (pY511)
Brand: PE Mouse Anti-Btk (pY551)/Itk (pY511)
SKU: 558129

Clone 24a/BTK (Y551) (RUO)

Analysis of Btk (pY551) in activated human B lymphoma cells. Ramos cells (ATCC CRL-1596) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE anti-Btk (pY551)/Itk (pY511). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

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Product Details

Brand: BD Phosflow™

Reactivity: Human (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: Phosphorylated Human Btk Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 20 µl

RRID: AB_397046

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
All other brands are trademarks of their respective owners.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Perm Buffer III RUO

Size 125 mL Cat No. 558050

Fixation Buffer RUO

Size 100 mL Cat No. 554655

558129 Rev. 4

Antibody Details

24a/BTK (Y551)

Bruton's tyrosine kinase (Btk) is a nonreceptor tyrosine kinase whose function is critical for proper B cell development and signaling. The activity of Btk is regulated by Src mediated phosphorylation of the kinase domain at tyrosine 551 (Y551). This event induces Btk kinase activity and subsequent autophosphorylation at Y223. Phosphorylated Btk then associates with the cell membrane via the interaction of the PH domain with phosphatidylinositol 3, 4, 5-triphosphate.

The Tec family kinase Itk plays a critical role in signal transduction downstream of the T cell antigen receptor and has been implicated in the activation of phospholipase C-γ1 (PLCγ1), a key regulator of calcium mobilization and extracellular signal-regulated kinase (ERK) activation. Itk is regulated by an activating transphosphorylation event in which Y511 in the kinase domain is phosphorylated by Lck.

The 24a/BTK (Y551) monoclonal antibody recognizes the Y551-phosphorylated form of human Btk and the Y511 phosphorylated form of human Itk.

558129 Rev. 4

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

558129 Rev.4

Citations & References

View product citations for antibody "558129" on CiteAb

Development References (3)

Mahajan S, Fargnoli J, Burkhardt AL, Kut SA, Saouaf SJ, Bolen JB. Src family protein tyrosine kinases induce autoactivation of Bruton's tyrosine kinase. Mol Cell Biol. 1995; 15:5304-5311. (Biology). View Reference
Rawlings DJ, Scharenberg AM, Park H, et al. Activation of BTK by a phosphorylation mechanism initiated by SRC family kinases. Science. 1996; 271:822-825. (Biology). View Reference
Wilcox HM, Berg LJ.. Itk phosphorylation sites are required for functional activity in primary T cells. J Biol Chem. 2003; 278:37112-37121. (Biology).

558129 Rev. 4

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.