Anti-BTK (phospho Y223) antibody (ab101233)
Key features and details
- Rabbit polyclonal to BTK (phospho Y223)
- Suitable for: WB, ELISA
- Reacts with: Human
- Isotype: IgG
Overview
-
Product name
Anti-BTK (phospho Y223) antibody
See all BTK primary antibodies -
Description
Rabbit polyclonal to BTK (phospho Y223) -
Host species
Rabbit -
Tested applications
Suitable for: WB, ELISAmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Cat, Dog, Pig, Chimpanzee, Macaque monkey, Orangutan
-
Immunogen
Synthetic peptide corresponding to Human BTK aa 200-300 (phospho Y223) conjugated to keyhole limpet haemocyanin.
-
Positive control
- This antibody gave a positive signal in Serum starved HeLa whole cell lysate
-
General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab101233 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 74 kDa (predicted molecular weight: 76 kDa). ELISA Use a concentration of 1 µg/ml. Target
-
Function
Plays a crucial role in B-cell ontogeny. Transiently phosphorylates GTF2I on tyrosine residues in response to B-cell receptor cross-linking. Required for the formation of functional ARID3A DNA-binding complexes. -
Involvement in disease
Defects in BTK are the cause of X-linked agammaglobulinemia (XLA) [MIM:300755]; also known as X-linked agammaglobulinemia type 1 (AGMX1) or immunodeficiency type 1 (IMD1). XLA is a humoral immunodeficiency disease which results in developmental defects in the maturation pathway of B-cells. Affected boys have normal levels of pre-B-cells in their bone marrow but virtually no circulating mature B-lymphocytes. This results in a lack of immunoglobulins of all classes and leads to recurrent bacterial infections like otitis, conjunctivitis, dermatitis, sinusitis in the first few years of life, or even some patients present overwhelming sepsis or meningitis, resulting in death in a few hours. Treatment in most cases is by infusion of intravenous immunoglobulin.
Defects in BTK may be the cause of X-linked hypogammaglobulinemia and isolated growth hormone deficiency (XLA-IGHD) [MIM:307200]; also known as agammaglobulinemia and isolated growth hormone deficiency or Fleisher syndrome or isolated growth hormone deficiency type 3 (IGHD3). In rare cases XLA is inherited together with isolated growth hormone deficiency (IGHD). -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
Contains 1 Btk-type zinc finger.
Contains 1 PH domain.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain. -
Post-translational
modificationsAutophosphorylated on Tyr-223 and Tyr-551. Phosphorylation of Tyr-223 may create a docking site for a SH2 containing protein. -
Cellular localization
Cytoplasm. Membrane. Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 695 Human
- Entrez Gene: 12229 Mouse
- Entrez Gene: 367901 Rat
- Omim: 300300 Human
- SwissProt: Q06187 Human
- SwissProt: P35991 Mouse
- Unigene: 159494 Human
- Unigene: 4475 Mouse
see all -
Alternative names
- Agammaglobulinaemia tyrosine kinase antibody
- AGMX 1 antibody
- AGMX1 antibody
see all
Images
-
Western blot - Anti-BTK (phospho Y223) antibody (ab101233)All lanes : Anti-BTK (phospho Y223) antibody (ab101233) at 1 µg/ml
Lane 1 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg
Lane 2 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg
Lane 3 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg with Immunising peptide at 1 µg/ml
Lane 4 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg with Immunising peptide at 1 µg/ml
Lane 5 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg with Non modified peptide at 1 µg/ml
Lane 6 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg with Non modified peptide at 1 µg/ml
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 76 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab101233 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
-
ELISA - Anti-BTK (phospho Y223) antibody (ab101233)ab101233 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
Datasheets and documents
References (1)
ab101233 has been referenced in 1 publication.
- Browning RL et al. Lenalidomide Induces Interleukin-21 Production by T Cells and Enhances IL21-Mediated Cytotoxicity in Chronic Lymphocytic Leukemia B Cells. Cancer Immunol Res 4:698-707 (2016). PubMed: 27287425
Images
-
Western blot - Anti-BTK (phospho Y223) antibody (ab101233)All lanes : Anti-BTK (phospho Y223) antibody (ab101233) at 1 µg/ml
Lane 1 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg
Lane 2 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg
Lane 3 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg with Immunising peptide at 1 µg/ml
Lane 4 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg with Immunising peptide at 1 µg/ml
Lane 5 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 10 µg with Non modified peptide at 1 µg/ml
Lane 6 : Hela (Human epithelial carcinoma cell line) Serum starved Whole Cell Lysate at 25 µg with Non modified peptide at 1 µg/ml
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 76 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab101233 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
-
ELISA - Anti-BTK (phospho Y223) antibody (ab101233)
ab101233 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
