Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD272/BTLA antibody [EPR20539] (ab212089)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20539] to CD272/BTLA
Suitable for: Flow Cyt, IP, WB, ICC/IF
Reacts with: Mouse

Product name

Anti-CD272/BTLA antibody [EPR20539]
See all CD272/BTLA primary antibodies
Description

Rabbit monoclonal [EPR20539] to CD272/BTLA
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A20 whole cell lysate; Mouse spleen and lymph node lysates. ICC/IF: A20 cells. Flow Cyt: A20 cells and mouse splenocytes. IP: Mouse spleen lysate.
General notes
This product was previously labelled as CD272

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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20539
Isotype

IgG
Research areas

Western blot - Anti-CD272/BTLA antibody [EPR20539] (ab212089)

Lane 1 : Anti-CD272/BTLA antibody [EPR20539] (ab212089) at 1/10000 dilution
Lanes 2-3 : Anti-CD272/BTLA antibody [EPR20539] (ab212089) at 1/1000 dilution

Lane 1 : A20 (mouse reticulum sarcoma cell line) whole cell lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Mouse lymph node 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: 33 kDa
Observed band size: 55-70 kDa
why is the actual band size different from the predicted?

Exposure time : Lane 1: 5 seconds; Lanes 2-3: 30 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-CD272/BTLA antibody [EPR20539] (ab212089)

Immunofluorescent analysis of 100% methanol fixed A20 (mouse reticulum sarcoma cell line) cells labeling CD272/BTLA with ab212089 at 1/100 dilution, followed by by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on A20 cell line.

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-CD272/BTLA antibody [EPR20539] (ab212089)

Flow cytometric analysis of mouse splenocytes labeling CD272/BTLA with ab212089 at 1/500 dilution (right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

Mouse splenocytes stained with CD3 and ab212089. Total viable cells were gated for the FC image. T cells (CD3+, upper population) show a moderate positive signal while B cells (lower right population in right panel) show stronger positive signal. The expression pattern is also reported in literature (PMID: 19949073).
Flow Cytometry - Anti-CD272/BTLA antibody [EPR20539] (ab212089)

Flow cytometric analysis of A20 (mouse reticulum sarcoma cell line) cell line labeling CD272/BTLA with ab212089 at 1/500 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) at 1/2000 dilution was used as the secondary antibody.

Total viable cells were gated for the FC image.
Immunoprecipitation - Anti-CD272/BTLA antibody [EPR20539] (ab212089)

CD272/BTLA was immunoprecipitated from 0.35 mg mouse spleen lysate with ab212089 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab212089 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution

Lane 1: Mouse spleen lysate 10 μg (Input).

Lane 2: ab212089 IP in mouse spleen lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab212089 in mouse spleen lysate.

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

Exposure time: 1 second.
Anti-CD272/BTLA antibody [EPR20539] (ab212089)

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