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AB277089

Human BTK ELISA Kit

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Human BTK ELISA Kit is a single-wash 90-min Simplestep used to quantify Human BTK with a sensitivity of 29 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

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

2 Images
Sandwich ELISA - Human BTK ELISA Kit (AB277089)
  • sELISA

Supplier Data

Sandwich ELISA - Human BTK ELISA Kit (AB277089)

Interpolated concentrations of native BTK in human 50 g/mL Raji and 100 g/mL Ramos cell extract load.

The concentrations of BTK were measured in duplicate and interpolated from the BTK standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean BTK concentration was determined to be 5,508.56 pg/mL in Raji cell extract and 5,046.75 pg/mL in Ramos cell extract.

Sandwich ELISA - Human BTK ELISA Kit (AB277089)
  • sELISA

Supplier Data

Sandwich ELISA - Human BTK ELISA Kit (AB277089)

Example of human BTK standard curve in 1X Cell Extraction Buffer PTR.

The BTK standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 29 pg/mL

Range

156.25 - 10000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human BTK ELISA kit (ab277089) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human BTK protein in human cell extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human BTK with 29 pg/mL sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

BTK, Bruton's tyrosine kinase, is critical for B-cell development. It sits at a major nexus for downstream signaling via phosphorylation cascades as well as phosphatidylinositol messaging. It is known to be responsible for oncogenic signaling that allows proliferation of B-cell leukemias. The antibodies in this kit were raised against the human full-length recombinant protein.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.99" }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.2" } ]
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Recovery

[ { "sample": "Cell Lysate", "range": "92 - 112 %", "average": "= 100" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab277089-1x96Tests", "assetComponentDetails": [ { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB277089-CMP04", "productcode":"" }, { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB277089-CMP05", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB277089-CMP11", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB277089-CMP12", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB277089-CMP06", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB277089-CMP10", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human BTK Lyophilized Recombinant Protein", "number":"AB277089-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human BTK Capture Antibody", "number":"AB277089-CMP01", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BI", "number":"AB277089-CMP08", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB277089-CMP07", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human BTK Detector Antibody", "number":"AB277089-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

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

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.

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.
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Product protocols

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Target data

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). Plays a role in STING1-mediated induction of type I interferon (IFN) response by phosphorylating DDX41 (PubMed : 25704810).
See full target information BTK
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