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Rabbit Recombinant Monoclonal BTK phospho Y551 antibody. Carrier free. Suitable for Flow Cyt (Intra), Dot, WB, I-ELISA, ICC/IF and reacts with Human samples.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (AB247285), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (AB247285), expandable thumbnail
  • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (AB247285), expandable thumbnail
  • Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (AB247285), expandable thumbnail
  • Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (AB247285), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow Cyt (Intra)IPFlow CytDotWBI-ELISAICC/IF
Human
Tested
Not recommended
Not recommended
Tested
Tested
Tested
Tested

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Target data

Function

Non-receptor tyrosine kinase indispensable for B lymphocyte development, differentiation and signaling (PubMed:19290921). Binding of antigen to the B-cell antigen receptor (BCR) triggers signaling that ultimately leads to B-cell activation (PubMed:19290921). After BCR engagement and activation at the plasma membrane, phosphorylates PLCG2 at several sites, igniting the downstream signaling pathway through calcium mobilization, followed by activation of the protein kinase C (PKC) family members (PubMed:11606584). PLCG2 phosphorylation is performed in close cooperation with the adapter protein B-cell linker protein BLNK (PubMed:11606584). BTK acts as a platform to bring together a diverse array of signaling proteins and is implicated in cytokine receptor signaling pathways (PubMed:16517732, PubMed:17932028). Plays an important role in the function of immune cells of innate as well as adaptive immunity, as a component of the Toll-like receptors (TLR) pathway (PubMed:16517732). The TLR pathway acts as a primary surveillance system for the detection of pathogens and are crucial to the activation of host defense (PubMed:16517732). Especially, is a critical molecule in regulating TLR9 activation in splenic B-cells (PubMed:16517732, PubMed:17932028). Within the TLR pathway, induces tyrosine phosphorylation of TIRAP which leads to TIRAP degradation (PubMed:16415872). BTK also plays a critical role in transcription regulation (PubMed:19290921). Induces the activity of NF-kappa-B, which is involved in regulating the expression of hundreds of genes (PubMed:19290921). BTK is involved on the signaling pathway linking TLR8 and TLR9 to NF-kappa-B (PubMed:19290921). Acts as an activator of NLRP3 inflammasome assembly by mediating phosphorylation of NLRP3 (PubMed:34554188). Transiently phosphorylates transcription factor GTF2I on tyrosine residues in response to BCR (PubMed:9012831). GTF2I then translocates to the nucleus to bind regulatory enhancer elements to modulate gene expression (PubMed:9012831). ARID3A and NFAT are other transcriptional target of BTK (PubMed:16738337). BTK is required for the formation of functional ARID3A DNA-binding complexes (PubMed:16738337). There is however no evidence that BTK itself binds directly to DNA (PubMed:16738337). BTK has a dual role in the regulation of apoptosis (PubMed:9751072).

Alternative names

AGMX1, ATK, BPK, BTK, Tyrosine-protein kinase BTK, Agammaglobulinemia tyrosine kinase, B-cell progenitor kinase, Bruton tyrosine kinase

Recommended products

Rabbit Recombinant Monoclonal BTK phospho Y551 antibody. Carrier free. Suitable for Flow Cyt (Intra), Dot, WB, I-ELISA, ICC/IF and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EP267Y
Purification technique
Affinity purification Protein A
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).

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab247285 is the carrier-free version of Anti-BTK (phospho Y551) antibody [EP267Y] ab40770.

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

BTK or Bruton's tyrosine kinase is an important enzyme in the body that functions mainly in signaling pathways within cells. It is known for its involvement in the development and activation of B cells and is expressed in cells of the hematopoietic system. The BTK protein has a molecular weight of about 76 kDa. Researchers can study BTK using tools like the BTK ELISA kit which helps measure its presence in biological samples. Furthermore there are specific anti-BTK antibodies which assist in the detection of BTK bands during analysis. BTK inhibitors like BTK C481S have been developed to regulate its activity.

Biological function summary

BTK plays an important role in the immune response by transmitting signals from the B cell receptor to the inside of the cell which promotes B cell maturation and survival. It operates as part of a larger signal transduction complex that includes other proteins and molecules. For accurate quantification of BTK expression a BTK sandwich ELISA kit may be utilized capturing and revealing the BTK protein's presence in sample preparations.

Pathways

BTK associates with both the B cell receptor signaling and the PI3K-Akt pathway. These pathways are critical for the proper functioning and proliferation of B cells. BTK interacts with proteins like PLCγ2 and BLNK in the signaling cascade highlighting its central role in transmitting extracellular signals to elicit appropriate cellular responses.

Associated diseases and disorders

BTK is closely linked to X-linked agammaglobulinemia (XLA) a genetic condition marked by an absence of mature B cells. This highlights the connection between BTK function and immune competence. Moreover aberrant BTK activity has been implicated in some types of B cell malignancies such as chronic lymphocytic leukemia (CLL). The BTK pathway's dysregulation in these disorders may involve interactions with proteins like SYK suggesting that targeting BTK could offer therapeutic benefits in managing such conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    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) (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

  • Flow Cytometry (Intracellular) - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

    Flow cytometric analysis of 4% paraformaldehyde fixed, 0.1% Tween-20 permeabilized Untreated control (Left) / Ramos cell treated with 1mM pervanadate for 30min (Middle) / Ramos cell treated with 1mM pervanadate for 30min, then incubated with Alkaline phosphatase overnight (Right) cells labelling BTK (phospho Y551) with Anti-BTK (phospho Y551) antibody [EP267Y] ab40770 at 1/2000 dilution (0.1µg) followed by a Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at a 1/5000 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    Blocking 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 (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

    Lanes 1 - 3: Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285) at 1/1000 dilution

    Lanes 1 - 3: Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770) at 1/1000 dilution

    Lane 1: Untreated Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg

    Lane 2: Ramos treated with 1mM pervanadate for 30 min whole cell lysate at 15 µg

    Lane 3: Ramos treated with 1mM pervanadate for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 76 kDa

    Observed band size: 75 kDa

    Exposure time: 7s

  • Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    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 Anti-BTK (phospho Y551) antibody [EP267Y] ab40770 at a dilution of 1/1000. Goat Anti-Rabbit IgG H&L (HRP) 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 (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

  • Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    ELISA using Anti-BTK (phospho Y551) antibody [EP267Y] 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 (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

  • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285), expandable thumbnail

    Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285)

    Blocking 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 (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770).

    Lanes 1 - 3: Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] - BSA and Azide free (ab247285) at 1/1000 dilution

    Lanes 1 - 3: Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (Anti-BTK (phospho Y551) antibody [EP267Y] ab40770) at 1/1000 dilution

    Lane 1: Untreated K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

    Lane 2: K-562 treated with 1mM pervanadate for 30 min whole cell lysate at 15 µg

    Lane 3: K-562 treated with 1mM pervanadate for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 76 kDa

    Observed band size: 75 kDa

    Exposure time: 180s

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Product protocols

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