PA5-20085

antibody from Invitrogen Antibodies

Targeting: BTK AGMX1, ATK, IMD1, PSCTK1, XLA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-20085

Provider

Invitrogen Antibodies

Product name

Btk Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

A suggested positive control is U937 cell lysate.

Concentration

1 mg/mL

References

Submitted references

Safety, Tolerability, Pharmacokinetics, Target Occupancy, and Concentration-QT Analysis of the Novel BTK Inhibitor Evobrutinib in Healthy Volunteers.

Becker A, Martin EC, Mitchell DY, Grenningloh R, Bender AT, Laurent J, Mackenzie H, Johne A
Clinical and translational science 2020 Mar;13(2):325-336

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of U937 cell lysate using a BTK polyclonal antibody (Product # PA5-20085) at (A) 0.5, (B) 1, and (C) 2 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BTK in U937 cell lysate with Btk Polyclonal Antibody (Product # PA5-20085) at (A) 0.5, (B) 1, and (C) 2 µg/mL.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Btk was performed using 70% confluent log phase Ramos cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with anti-Btk Polyclonal Antibody (Product # PA5-20085) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma Membrane and Cytoplasmic localization. Panel e represents negative control, Jurkat cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry of BTK in Daudi cells with Btk Polyclonal Antibody (Product # PA5-20085) at 10 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of BTK in Daudi cells with Btk Polyclonal Antibody (Product # PA5-20085) at 10 µg/mL.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of BTK in mouse brain tissue with Btk Polyclonal Antibody (Product # PA5-20085) at 20 µg/mL. Red: BTK Blue: DAPI staining

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of BTK in mouse brain tissue with Btk Polyclonal Antibody (Product # PA5-20085) at 20 µg/mL. Red: BTK Blue: DAPI staining

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of BTK in mouse brain tissue with Btk Polyclonal Antibody (Product # PA5-20085) at 5 µg/mL.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 Arithmetic mean (SD) of % Bruton's tyrosine kinase (BTK) receptor occupancy* vs. time after administration of evobrutinib for ( a ) single dosing and ( b ) multiple dosing. *% BTK occupancy = (1-(BTK F /BTK T )/(BTK F pre-dose/BTK T pre-dose))*100. BTK T , total Bruton's tyrosine kinase; BTK F , free Bruton's tyrosine kinase.