Anti-BTK antibody [EPR20445] (ab208937)

Anti-BTK antibody [EPR20445] (ab208937) is a rabbit monoclonal antibody that is used to detect BTK in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] (AB208937), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Tested	Tested	Not recommended
Rat	Expected	Tested	Expected	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -
Species Mouse	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/600	Notes -
Species Mouse	Dilution info 1/600	Notes -
Species Rat	Dilution info 1/600	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information BTK

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20445
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-BTK antibody [EPR20445] (ab208937) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of BTK?

Anti-BTK [EPR20445] (ab208937) specifically detects a band for BTK (UniProt: Q06187) at a molecular weight of 76kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR20445] also available for your convenience:
ab208937, Carrier free - ab227812, APC - ab317755, Alexa Fluor® 647 - ab319012, FITC - ab322295

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

What are the advantages of a recombinant monoclonal antibody?
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.

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

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14 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] (ab208937)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining on lymphoid nodule in human colon (PMID: 25433814). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-BTK antibody [EPR20445] (ab208937)
Immunofluorescent analysis of 100% methanol-fixed Ramos (human Burkitt's lymphoma cell line) and Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling BTK with ab208937 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on Ramos cell line.
Negative control: Jurkat cell line (PMID: 24759210).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking/Dilution buffer: 5% NFDM/TBST.
The Jurkat T cell line is negative for BTK (PMID 24759210).
This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
Lane 1: Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2: NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 76 kDa
Exposure time: 32s
Flow Cytometry (Intracellular) - Anti-BTK antibody [EPR20445] (ab208937)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and Ramos (human Burkitt's lymphoma cell line) (right panel) cell lines labeling BTK with ab208937 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: Jurkat cell line (PMID: 24759210).
Immunoprecipitation - Anti-BTK antibody [EPR20445] (ab208937)
BTK was immunoprecipitated from 0.35 mg of Ramos (human Burkitt's lymphoma cell line) whole cell lysate with ab208937 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208937 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Ramos whole cell lysate 10 μg (Input).
Lane 2: ab208937 IP in Ramos whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab208937 in Ramos whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
All lanes: Immunoprecipitation - Anti-BTK antibody [EPR20445] (ab208937)
Predicted band size: 76 kDa
Observed band size: 76 kDa
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking/Dilution buffer: 5% NFDM/TBST.
This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
All lanes: Human lymph node lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 76 kDa
Exposure time: 3min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] (ab208937)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining in mantle zone and weaker cytoplasmic staining in the germinal center of human tonsil (PMID: 25433814). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
Lane 1: THP-1 (human monocytic leukemia cell) whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 3: NR8383 (rat lung macrophage (alveolar)) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDa
Exposure time: 37s
Flow Cytometry (Intracellular) - Anti-BTK antibody [EPR20445] (ab208937)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized NR8383 (rat alveolar macrophage) cell lines labeling BTK with ab208937 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used as the secondary antibody at 1/5000 dilution.
Flow Cytometry (Intracellular) - Anti-BTK antibody [EPR20445] (ab208937)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell lines labeling BTK with ab208937 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used as the secondary antibody at 1/5000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-BTK antibody [EPR20445] (ab208937)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling BTK with ab208937 at 1/250 (8.6 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 (2 ug/ml) dilution (green). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^®
Confocal image showing cytoplasmic staining in RAW 264.7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 15s
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
Lane 1: THP-1 (human monocytic leukemia cell) whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 3: NR8383 (rat lung macrophage (alveolar)) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDa
Exposure time: 37s
Western blot - Anti-BTK antibody [EPR20445] (ab208937)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
All lanes: Western blot - Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution
Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDa
Exposure time: 15s