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<strong> Figure 1 WB Validation in Human Cell Lines </strong><br>Loading: 15 μg of lysate Antibodies: BTK, 3395, 0.5 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 WB Validation in Mouse Tissues and Cells</strong><br>Loading: 15 μg of lysate Antibodies: BTK, 3395, 0.5 μ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 WB Validation in Rat Spleen</strong><br>Loading: 15 μg of lysate Antibodies: BTK, 3395, 0.5 μ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunofluorescence Validation of BTK In THP-1 Cells</strong><br>Immunofluorescent analysis of PFA-fixed THP-1 cells labeling BTK with 3395 at 20 ?g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 5 Immunofluorescence Validation of BTK in Mouse Brain</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain tissue labeling BTK with 3395 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
<strong>Figure 6 Immunohistochemistry Validation of BTK in Human Spleen</strong><br> Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-BTK antibody (3395) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunohistochemistry Validation of BTK in Mouse Spleen</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-BTK antibody (3395) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 8 Immunohistochemistry Validation of BTK in Rat spleen</strong><br> Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-BTK antibody (3395) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 9 Immunohistochemistry Validation of BTK in Mouse Brain</strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-BTK antibody (3395) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Figure 1 WB Validation in Human Cell Lines
Loading: 15 μg of lysate Antibodies: BTK, 3395, 0.5 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

BTK Antibody

CATALOG NUMBER: 3395

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB, ICC/IF, IHC-P/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Chicken: (100%)
Immunogen:
Anti-BTK antibody (3395) was raised against a peptide corresponding to 16 amino acid synthetic peptide near the amino terminus of human BTK.

The immunogen is located within amino acids 20-70 of BTK.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB, ICC/IF, IHC-P/IF
Application Note:
WB: 0.5-2 μg/mL; ICC/IF: 10-20 μg/mL; IHC-P/IF: 10-20 μg/mL; IHC-P: 1-5 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples; Immunofluorescence in human and mouse samples. All other applications and species not yet tested.
Specificity:
Multiple isoforms of BTK are known to exist.
Positive Control 1:
Cat. No. 1406 - Mouse Spleen Tissue Lysate
Positive Control 2:
Cat. No. 1466 - Rat Spleen Tissue Lysate
Predicted Molecular Weight:
Predicted: 76 kD

Observed: 72 kD
Purification:
BTK Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
BTK Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
BTK antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
BTK
Additional Names:
BTK Antibody: AT, ATK, BPK, XLA, IMD1, AGMX1, PSCTK1, Tyrosine-protein kinase BTK, Agammaglobulinemia tyrosine kinase
Protein Accession Number:
Q06187
Protein Gi Number:
547759
Ncbi Gene Id Number:
695
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
BTK Antibody: Bruton's tyrosine kinase (BTK) was initially identified as a member of the src family for protein-tyrosine kinases that was involved in X-linked agamma-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.
Background Reference 1:
Vetrie D, Vorechovsky I, Sideras P, et al. The gene involved in X-linked aγglobulinaemia is a member of the src family of tyrosine kinases. Nature 1993; 361:226-33.
Background Reference 2:
Kawakami Y, Kitaura J, Hata D, et al. Functions of Bruton's tyrosine kinase in mast and B cells. J. Leukoc. Biol. 1999; 65(3):286-90.
Background Reference 3:
Jefferies CA, Doyla S, Brunner C, et al. Bruton's tyrosine kinase is a Toll/Interleukin-1 receptor domain-binding protein that participates in nuclear factor kB activation by Toll-like receptor 4. J. Biol. Chem. 2003; 278:26258-26264.
Background Reference 4:
Akira S and Takeda K. Toll-like receptor Signalling. Nat. Rev. Immunol. 2004; 4:499-511.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

3395

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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