BTK Recombinant Rabbit Monoclonal Antibody [SN69-07] - BSA and Azide free
Catalog# HA750252
BTK Recombinant Rabbit Monoclonal Antibody [SN69-07] - BSA and Azide free
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WB
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IHC-P
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IP
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IF-Cell
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Human
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Mouse
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ET1611-4
含抗保成分
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unconjugated
Overview
Product Name
BTK Recombinant Rabbit Monoclonal Antibody [SN69-07] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human BTK aa 610-659 / 659.
Species Reactivity
Human, Mouse
Validated Applications
WB, IHC-P, IP, IF-Cell
Molecular Weight
Predicted band size: 76 kDa
Positive Control
Ramos cell lysate, Raji cell lysate, THP-1 cell lysate, Daudi cell lysate, RAW264.7 cell lysate, human tonsil tissue.
Conjugation
unconjugated
Clone Number
SN69-07
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IHC-P
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1:50-1:100
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IP
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Use at an assay dependent concentration.
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IF-Cell
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1:100
Target
Function
The Tec family of non-receptor tyrosine kinases is composed of six proteins designated Tec, Emt (also known as Itk or Tsk), Btk (previously known as Atk, BPK or Emb), Bmx, Txk (also known as Rlk) and Dsrc28C. All members of the family contain SH3 and SH2 domains and, with the exception of Txk and Dsrc28C, also contain a pleckstrin homology (PH) and a Tec homology (TH) domain in their amino termini. Four alternatively spliced forms of Tec are found to be expressed broadly in cells of hematopoietic lineage and hepatocytes. The Emt gene product associates with CD28 and becomes activated subsequent to CD28 ligation. Btk is necessary for proper B cell development, and mutations in the gene encoding Btk have been associated with families suffering from X-linked agammaglobulinemia, also referred to as Bruton's disease. The Bmx protein shares a high degree of homology with Btk and seems to be expressed at highest levels in the heart. Txk expression is T cell-specific, while expression of the Drosophila Tec homolog, Dsrc28C, is developmentally regulated.
Background References
1. Landon AL et al. MNKs act as a regulatory switch for eIF4E1 and eIF4E3 driven mRNA translation in DLBCL. Nat Commun 5:5413 (2014).
2. Bogusz AM et al. Quantitative Immunofluorescence Reveals the Signature of Active B-cell Receptor Signaling in Diffuse Large B-cell Lymphoma. Clin Cancer Res 18:6122-35 (2012).
Sequence Similarity
Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
Tissue Specificity
Predominantly expressed in B-lymphocytes.
Post-translational Modification
Following B-cell receptor (BCR) engagement, translocates to the plasma membrane where it gets phosphorylated at Tyr-551 by LYN and SYK. Phosphorylation at Tyr-551 is followed by autophosphorylation of Tyr-223 which may create a docking site for a SH2 containing protein. Phosphorylation at Ser-180 by PRKCB, leads in translocation of BTK back to the cytoplasmic fraction. Phosphorylation at Ser-21 and Ser-115 creates a binding site for PIN1 at these Ser-Pro motifs, and promotes it's recruitment.
Subcellular Location
Cell membrane, Cytoplasm, Membrane, Nucleus.
Synonyms
Agammaglobulinaemia tyrosine kinase antibody
AGMX 1 antibody
AGMX1 antibody
AT antibody
ATK antibody
B cell progenitor kinase antibody
B-cell progenitor kinase antibody
BPK antibody
Bruton agammaglobulinemia tyrosine kinase antibody
Bruton tyrosine kinase antibody
ExpandImages
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Western blot analysis of BTK on different lysates with Rabbit anti-BTK antibody (HA750252) at 1/5,000 dilution.
Lane 1: Ramos cell lysate
Lane 2: Jurkat cell lysate (negative)
Lane 3: Raji cell lysate
Lane 4: THP-1 cell lysate
Lane 5: Daudi cell lysate
Lane 6: RAW264.7 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 76 kDa
Observed band size: 72 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750252) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockout (KO)
All lanes: Western blot analysis of BTK with anti-BTK antibody [SN69-07] (HA750252) at 1/1,000 dilution.
Lane 1: Wild-type U2932 whole cell lysate.
Lane 2: BTK knockout U2932 whole cell lysate.
ET1611-4 was shown to specifically react with BTK in wild-type U2932 cells. No band was observed when BTK knockout sample was tested. Wild-type and BTK knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-BTK antibody (ET1611-4, 1/1,000) and Anti-Vinculin antibody (ET1705-94, 1/5,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China). -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-BTK antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750252, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Application: Immunocytochemistry (IF-cell)
Species: Mouse
Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA750252, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
BTK Recombinant Rabbit Monoclonal Antibody [SN69-07]
Application: WB,IHC-P,IP,IF-Cell
Reactivity: Human,Mouse
Conjugate: unconjugated
