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 nameAnti-CD272/BTLA antibody [EPR20539]
See all CD272/BTLA primary antibodies
DescriptionRabbit monoclonal [EPR20539] to CD272/BTLA
Tested applicationsSuitable for: Flow Cyt, IP, WB, ICC/IFmore details
Species reactivityReacts with: Mouse
Recombinant fragment within Mouse CD272/BTLA aa 1-200. The exact sequence is proprietary.
Database link: Q7TSA3
- 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.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped 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 bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab212089 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 55-70 kDa (predicted molecular weight: 33 kDa).|
FunctionLymphocyte inhibitory receptor which inhibits lymphocytes during immune response.
Sequence similaritiesContains 1 Ig-like V-type (immunoglobulin-like) domain.
modificationsPhosphorylated on Tyr residues by TNFRSF14 and by antigen receptors cross-linking, both inducing association with PTPN6 and PTPN11.
- Information by UniProt
- B and T lymphocyte associated protein antibody
- B and T lymphocyte attenuator antibody
- B and T lymphocyte associated antibody
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.
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.
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 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 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.
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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab212089 has not yet been referenced specifically in any publications.