Immunohistochemistry analysis of Phospho-BTK [pTyr551] showing staining in the cytoplasm of paraffin-embedded human spleen tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-BTK [pTyr551] polyclonal antibody (441355G) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human BTK that contains tyrosine 551.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Bruton and quote;s tyrosine kinase (BTK) was initially identified as a member of the src family for protein-tyrosine kinases that was involved in X-linked a-gamma-globulinaemia, and has since been shown to involved in a number of signaling pathways in hemapoietic lineage. It has recently been shown to interact with members of the toll-like receptor (TLR) family such as TLR4, 6, 8, and 9. The TLRs are critical molecules in both the innate and adaptive immunity and can recognize diverse microbial pathogens. BTK has also been shown to interact with key proteins involved in TLR4 signal transduction such as MyD88, TIRAP, and IRAK, but not TRAF-6, suggesting that BTK is involved in lipopolysaccharide signal transduction.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.