Antibodies, Proteins, Kits and Reagents for Life Science

278 083 ₸ Product size

100 µl 278 083 ₸

Order now and get it on Tuesday March 09, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20445] to BTK
Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt
Reacts with: Human

Product name

Anti-BTK antibody [EPR20445]
See all BTK primary antibodies
Description

Rabbit monoclonal [EPR20445] to BTK
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Ramos and NAMALWA whole cell lysates; Human lymph node lysate. IHC-P: Human tonsil and colon tissues. ICC/IF: Ramos cells. Flow Cyt: Ramos cells. IP: Ramos whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20445
Isotype

IgG
Research areas

Western blot - Anti-BTK antibody [EPR20445] (ab208937)

All lanes : Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution

Lane 1 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 76 kDa
Observed band size: 76 kDa

Exposure time: 32 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The Jurkat T cell line is negative for BTK (PMID 24759210).

This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.
Western blot - Anti-BTK antibody [EPR20445] (ab208937)

Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution + Human lymph node lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 76 kDa
Observed band size: 76 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] (ab208937)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining in mantle zone and weaker cytoplasmic staining in the germinal center of human tonsil (PMID: 25433814). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] (ab208937)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining on lymphoid nodule in human colon (PMID: 25433814). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-BTK antibody [EPR20445] (ab208937)

Immunofluorescent analysis of 100% methanol-fixed Ramos (human Burkitt's lymphoma cell line) and Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling BTK with ab208937 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on Ramos cell line.

Negative control: Jurkat cell line (PMID: 24759210).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow Cytometry - Anti-BTK antibody [EPR20445] (ab208937)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and Ramos (human Burkitt's lymphoma cell line) (right panel) cell lines labeling BTK with ab208937 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control: Jurkat cell line (PMID: 24759210).
Immunoprecipitation - Anti-BTK antibody [EPR20445] (ab208937)

BTK was immunoprecipitated from 0.35 mg of Ramos (human Burkitt's lymphoma cell line) whole cell lysate with ab208937 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208937 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Ramos whole cell lysate 10 µg (Input).

Lane 2: ab208937 IP in Ramos whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208937 in Ramos whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Anti-BTK antibody [EPR20445] (ab208937)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com