Recombinant Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)
![Western blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)](https://www.abcam.com/ps/products/68/ab68217/Images/ab68217-5-ab68217WB1.jpg "Western blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP420Y] to BTK (phospho Y223)
- Suitable for: WB, IP, Dot blot
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-BTK (phospho Y223) antibody [EP420Y]
See all BTK primary antibodies -
Description
Rabbit monoclonal [EP420Y] to BTK (phospho Y223) -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Dot blotmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab214589) -
Positive control
- WB: K562 and Ramos cell lysates treated with pervanadate. IP: Ramos cell lysates treated with pervanadate. Dot Blot: BTK (phospho Y223) phospho peptide.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP420Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68217 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 |
1/2000 - 1/10000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).Can be blocked with BTK peptide (ab214589).
For unpurified, use 1/10000 - 1/50000. |
|
| IP |
1/20.
For unpurified, use 1/40. |
|
| Dot blot |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
WB
1/2000 - 1/10000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).Can be blocked with BTK peptide (ab214589). For unpurified, use 1/10000 - 1/50000. |
|
IP
1/20. For unpurified, use 1/40. |
|
Dot blot
Use at an assay dependent concentration. |
Target
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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
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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
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Alternative names
- Agammaglobulinaemia tyrosine kinase antibody
- AGMX 1 antibody
- AGMX1 antibody
see all
Images
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Western blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)All lanes : Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217) at 1/1000 dilution
Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate
Lane 3 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate. Then the membrane was incubated with alkaline phosphatase.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDaBlocking and dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)](https://www.abcam.com/ps/products/68/ab68217/Images/ab68217-239340-68217-IP.jpg)
Immunoprecipitation - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)ab68217 (purified) at 1/50 immunoprecipitating BTK (phospho Y223) in Ramos cells (Lane 1) treated with 1 mM Pervanadate for 30 min. For western blotting, a HRP-conjugated anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Dot Blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)](https://www.abcam.com/ps/products/68/ab68217/Images/ab68217-4-ab68217Dot1.jpg)
Dot Blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)Dot blot analysis of BTK (phospho Y223) phospho peptide (Lane 1) and BTK phospho peptide (Lane 2) labeling BTK (phospho Y223) with ab68217 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.
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![Western blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)](https://www.abcam.com/ps/products/68/ab68217/Images/ab68217-239337-68217-WB.jpg)
Western blot - Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)All lanes : Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217) at 1/10000 dilution (purified)
Lane 1 : Untreated K562 cell lysate
Lane 2 : K562 cells treated with pervanadate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDa -
![Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)](https://www.abcam.com/ps/products/68/ab68217/Images/ab68217-6-benefits-of-recombinant-antibodies.png)
Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (19)
ab68217 has been referenced in 19 publications.
- Chen T et al. The effect of geniposide on chronic unpredictable mild stress-induced depressive mice through BTK/TLR4/NF-?B and BDNF/TrkB signaling pathways. Phytother Res 35:932-945 (2021). PubMed: 33164233
- Eisenmann ED et al. Intentional Modulation of Ibrutinib Pharmacokinetics through CYP3A Inhibition. Cancer Res Commun 1:79-89 (2021). PubMed: 34950932
- Sheng L et al. Overexpression of Fc?RIIB regulates downstream protein phosphorylation and suppresses B cell activation to ameliorate systemic lupus erythematosus. Int J Mol Med 46:1409-1422 (2020). PubMed: 32945349
- Liu S et al. The protective effect of Geniposide on diabetic cognitive impairment through BTK/TLR4/NF-?B pathway. Psychopharmacology (Berl) 237:465-477 (2020). PubMed: 31811349
- Kim E et al. The BET inhibitor GS-5829 targets chronic lymphocytic leukemia cells and their supportive microenvironment. Leukemia 34:1588-1598 (2020). PubMed: 31862959