Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 720101
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Product Specifications

Immunogen

Phosphopeptide containing the human BTK Y223 site

Specificity

Detects human Phospho-BTK (Y223) in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Phospho-BTK (Y223) Antibody

Detection of Human Phospho-BTK (Y223) antibody by Western Blot.

Detection of Human Phospho-BTK (Y223) by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line untreated (-) or treated (+) with 1 mM Pervanadate for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-BTK (Y223) Monoclonal Antibody (Catalog # MAB7205) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-BTK (Y223) at approximately 80 kDa (as indicated). For additional reference, duplicate samples were probed with 0.05 µg/mL of Mouse Anti-Human BTK Monoclonal Antibody (lower panel, Catalog # MAB5807) This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-BTK (Y223) antibody in Raji Human Cell Line by Immunocytochemistry (ICC).

Phospho-BTK (Y223) in Raji Human Cell Line.

BTK phosphorylated at Y223 was detected in immersion fixed Raji human Burkitt's lymphoma cell line with (upper panel) and without (lower panel) pervanadate stimulation using Mouse Anti-Human Phospho-BTK (Y223) Monoclonal Antibody (Catalog # MAB7205) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human Phospho-BTK (Y223) Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed Raji human Burkitt's lymphoma cell line with and without pervanadate stimulation

Western Blot

1 µg/mL
Sample: Raji human Burkitt's lymphoma cell line treated with Pervanadate

Reviewed Applications

Read 2 reviews rated 5 using MAB7205 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 PLC gamma by BTK. The autophosphorylation of Tyr223 facilitates the interaction of BTK with activated Syk. The sequence used as a peptide immunogen is identical in human, 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 (Y223) Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Phospho-BTK (Y223) Antibody

For research use only

Customer Reviews for Human Phospho-BTK (Y223) Antibody (2)

5 out of 5
2 Customer Ratings
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1 Stars
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  • Human Phospho-BTK (Y223) Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HEK293 cells
    Species: Human
    Verified Customer | Posted 12/27/2021
    Human Phospho-BTK (Y223) Antibody MAB7205
  • Human Phospho-BTK (Y223) 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 (Y223) Antibody MAB7205

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Protocols

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