Human phospho-BTK (Y551) Antibody (797837)

R&D Systems | Catalog # MAB7659

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 797837
View all BTK Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human BTK Y551 site

Specificity

Detects human BTK when phosphorylated at Y551.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human phospho-BTK (Y551) Antibody (797837)

Detection of Human Phospho-BTK (Y551) antibody by Western Blot.

Detection of Human Phospho-BTK (Y551) by Western Blot.

Western blot shows lysates of RL human non-Hodgkin's lymphoma B cell line and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 0.1 mM Pervanadate (PV) for 10 minutes and 12 µg/mL Goat Anti-Human IgM µ Chain Antigen Affinity-purified Polyclonal Antibody (Catalog # G-105-C) for 2 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-BTK (Y551) Monoclonal Antibody (Catalog # MAB7659) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-BTK (Y551) at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-BTK (Y551) antibody by Simple WesternTM.

Detection of Human Phospho-BTK (Y551) by Simple WesternTM.

Simple Western lane view shows lysates of Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with ultraviolet light (UV), loaded at 0.2 mg/mL. A specific band was detected for Phospho-BTK (Y551) at approximately 89 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Phospho-BTK (Y551) Monoclonal Antibody (Catalog # MAB7659). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Phospho-BTK (Y551) by Western Blot

Detection of Phospho-BTK (Y551) by Western Blot

Antigen-triggered phosphorylation of BTK and PLC gamma in BMMCs is independent of PI3K. (a) WT BMMCs were pre-treated for 20 min with 100 nM Wortmannin (WM) or vehicle (DMSO) and were then left untreated (con) or stimulated with the indicated concentrations of Ag (DNP-HSA [DNP]) for 1 and 5 min. Btk−/− BMMCs were pre-treated with DMSO and stimulated accordingly. Whole-cell lysates were subjected to Western Blot analysis with antibodies against P-BTK (Tyr223), P-PKB (Ser473), and p85 (loading control). (b) WT BMMCs were pre-treated for 20 min with 100 nM WM or vehicle (DMSO) and were then left untreated (con) or stimulated with Ag (20 ng/ml DNP-HSA) for the indicated times. Btk−/− BMMCs were pre-treated with DMSO and stimulated accordingly. Whole-cell lysates were subjected to Western Blot analysis with antibodies against P-BTK (Tyr551), BTK, P-PKB (Ser473), and p85 (loading control). (c) WT and Btk−/− BMMCs were pre-treated for 20 min with 100 nM WM or vehicle (DMSO) and were then left untreated (con) or stimulated with the indicated concentrations of Ag for 1 and 5 min. Whole-cell lysates were analysed by immunoblotting against P-PLC gamma 1 (Tyr783), PLC gamma 1, P-PKB (Ser473), and p85 (loading control). Densitometry of P-PLC gamma 1 with reference to PLC gamma 1 was performed using ImageJ software (NIH, USA) and normalised to WT cells pretreated with DMSO and stimulated with 1 ng/ml Ag (1 min). Comparable results were obtained in at least three experiments with different BMMC cultures. Fine vertical lines were inserted to allow for better discrimination of cells/conditions. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30341350), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human phospho-BTK (Y551) Antibody (797837)

Application
Recommended Usage

Simple Western

20 µg/mL
Sample: Ramos human Burkitt's lymphoma cell line treated with ultraviolet light (UV)

Western Blot

1 µg/mL
Sample: RL human non-Hodgkin's lymphoma B cell line and Ramos human Burkitt's lymphoma cell line treated (+) Pervanadate (PV) and Goat Anti-Human IgM μ Chain Antigen Affinity-purified Polyclonal Antibody (Catalog # G-105-C) for 2 minutes

Reviewed Applications

Read 1 review rated 5 using MAB7659 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BTK

Bruton tyrosine kinase (Btk) is a 75 kDa cytoplasmic protein tyrosine kinase that is widely expressed in hematopoietic cells. Btk is required for B cell receptor signaling and B cell development. Defects in Btk result in X-linked agammaglobulinemia which is characterized by a severely decreased level of circulating antibodies. Like other Tec family kinases, Btk contains a Pleckstrin homology domain, a Tec homology domain, an SH3 domain, an SH2 domain, and a protein kinase domain. Association of the Btk SH2 domain with the B cell linker protein (BLNK) is required for the activation of PLCg by Btk. Btk can be activated by Lyn or Syk phosphorylation of Tyr551 within the catalytic domain. Within aa 546 - 556, human Btk shares 100% aa sequence identity with mouse and rat Btk.

Long Name

Bruton Agammaglobulinemia Tyrosine Kinase

Alternate Names

AGMX1, IMD1, PSCTK1, XLA

Entrez Gene IDs

695 (Human)

Gene Symbol

BTK

Additional BTK Products

Product Documents for Human phospho-BTK (Y551) Antibody (797837)

Certificate of Analysis

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Product Specific Notices for Human phospho-BTK (Y551) Antibody (797837)

For research use only

Citations for Human phospho-BTK (Y551) Antibody (797837)

Customer Reviews for Human phospho-BTK (Y551) Antibody (797837) (1)

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  • Human Phospho-BTK (Y551) Antibody
    Name: Rachel Welt
    Application: ELISA
    Sample Tested: B cell lymphoma
    Species: Human
    Verified Customer | Posted 01/08/2020
    This product was used as a capture antibody in an ELISA assay to test for BTK phosphorylation. Worked great!
    Human phospho-BTK (Y551) Antibody (797837) MAB7659

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