Recombinant Anti-LAG-3 antibody [EPR20294-77] (ab209238)
![Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)](https://www.abcam.com/ps/products/209/ab209238/Images/ab209238-293409-anti-lymphocyte-activation-gene-3-antibody-epr20294-77-immunofluorescence.jpg "Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20294-77] to LAG-3
- Suitable for: WB, ICC/IF, IP
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-LAG-3 antibody [EPR20294-77]
See all LAG-3 primary antibodies -
Description
Rabbit monoclonal [EPR20294-77] to LAG-3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus; mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours, whole cell lysate. ICC/IF: Mouse splenocytes. IP: Mouse splenocytes treated with 5 µg/ml concanavalin A for 72 hours, whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20294-77 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab209238 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 54, 70 kDa (predicted molecular weight: 57 kDa).
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| ICC/IF |
1/100.
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| IP |
1/30.
|
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 54, 70 kDa (predicted molecular weight: 57 kDa). |
|
ICC/IF
1/100. |
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IP
1/30. |
Target
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Function
Involved in lymphocyte activation. Binds to HLA class-II antigens. -
Tissue specificity
On cell surface of activated NK and T-lymphocytes. -
Sequence similarities
Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 16768 Mouse
- SwissProt: Q61790 Mouse
- Unigene: 4528 Mouse
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Alternative names
- CD223 antibody
- CD223 antigen antibody
- FDC protein antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized untreated and concanavalin A (2.5 μg/ml for 72 hours) treated mouse splenocytes labeling LAG-3 with ab209238 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 increased membranous staining following treatment of mouse splenocytes with concanavalin A (2.5 μg/ml for 72 hours) (PMID 20391435; PMID 26318293; PMID 11527700; PMID 19631993).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution
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![Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)](https://www.abcam.com/ps/products/209/ab209238/Images/ab209238-293405-anti-lymphocyte-activation-gene-3-antibody-epr20294-77-western-blot.jpg)
Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/10000 dilution + Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus, 10 ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution Buffer: 5% NFDM/TBST
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![Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)](https://www.abcam.com/ps/products/209/ab209238/Images/ab209238-293407-anti-lymphocyte-activation-gene-3-antibody-epr20294-77-western-blot.jpg)
Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)All lanes : Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/1000 dilution
Lane 1 : Untreated mouse splenocytes whole cell lysate
Lane 2 : Mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 54, 70 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Dilution Buffer: 5% NFDM/TBST
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
LAG3 expression can be elevated in activated splenocytes (PMID 19631993).
The molecular mass observed is consistent with the literature (PMID 15971272; PMID 15557174).
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![Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)](https://www.abcam.com/ps/products/209/ab209238/Images/ab209238-293425-anti-lymphocyte-activation-gene-3-antibody-epr20294-77-immunoprecipitation.jpg)
Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)LAG-3 was immunoprecipitated from 0.35 mg of mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours) whole cell lysate with ab209238 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209238 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: Mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate 10μg (Input).
Lane 2: ab209238 IP in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209238 in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 1 second.
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![Anti-LAG-3 antibody [EPR20294-77] (ab209238)](https://www.abcam.com/ps/products/209/ab209238/Images/ab209238-5-benefits-of-recombinant-antibodies.png)
Anti-LAG-3 antibody [EPR20294-77] (ab209238)
Protocols
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab209238 has been referenced in 1 publication.
- Wingrove E et al. Transcriptomic Hallmarks of Tumor Plasticity and Stromal Interactions in Brain Metastasis. Cell Rep 27:1277-1292.e7 (2019). PubMed: 31018140
