Recombinant Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP267Y] to BTK (phospho Y551) - BSA and Azide free
- Suitable for: WB, Dot blot, Indirect ELISA, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free
See all BTK primary antibodies -
Description
Rabbit monoclonal [EP267Y] to BTK (phospho Y551) - BSA and Azide free -
Host species
Rabbit -
Specificity
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
ab40770 also reacts with family proteins including BMX (phospho Y566), ITK (phospho Y512), TEC (phospho Y519) and TXK (phospho Y420).
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Tested applications
Suitable for: WB, Dot blot, Indirect ELISA, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Ramos treated with 1mM pervanadate whole cell lysates, K-562 treated with 1mM pervanadate whole cell lysate ICC/IF: Ramos cells
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General notes
ab247285 is the carrier-free version of ab40770.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP267Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247285 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 |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 74 kDa (predicted molecular weight: 74 kDa).
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Dot blot |
1/1000.
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Indirect ELISA |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 74 kDa (predicted molecular weight: 74 kDa). |
Dot blot
1/1000. |
Indirect ELISA
Use at an assay dependent concentration. |
ICC/IF
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
- Omim: 300300 Human
- SwissProt: Q06187 Human
- Unigene: 159494 Human
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Alternative names
- Agammaglobulinaemia tyrosine kinase antibody
- AGMX 1 antibody
- AGMX1 antibody
see all
Images
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ELISA using ab40770 at varying antibody concentrations and antigen concentration of 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody. Substrate solution was p-nitrophenyl phosphate (PNPP).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40770).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling BTK with primary antibody anti- BTK (phospho Y551) (ab40770) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing increased cytoplasmic staining in Ramos cells treated with pervanadate (1 mM) for 30 min and then the signal decreased after Alkaline Phosphatase treatment 37℃ for 1 hour. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).
The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40770).
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Dot blot analysis of Human BTK non-phospho peptide (lane 1), Human BTK Y551 phospho peptide (lane 2), Human BMX non-phospho peptide (lane 3), Human BMX Y566 phospho peptide (lane 4), Human ITK non-phospho peptide (lane 5), Human ITK Y512 phospho peptide (lane 6), Human TEC non-phospho peptide (lane 7), Human TEC Y519 phospho peptide (lane 8), Human TXK non-phospho peptide (lane 9) and Human TXK Y420 phospho peptide (lane 10) with ab40770 at a dilution of 1/1000. ab97051 Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/20,000.
Blocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBST
Exposure time: 180 seconds
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40770).
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All lanes : Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770) at 1/1000 dilution
Lane 1 : Untreated K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : K-562 treated with 1mM pervanadate for 30 min whole cell lysate
Lane 3 : K-562 treated with 1mM pervanadate for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 74 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40770).
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All lanes : Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770) at 1/1000 dilution
Lane 1 : Untreated 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 treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 74 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 7 secondsBlocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40770).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247285 has not yet been referenced specifically in any publications.