Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸ 10 µl 120 614 ₸

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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

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 & Species

Application	Species
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: 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.
General notes
This product was previously labelled as Lymphocyte Activation Gene 3.
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, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20294-77
Isotype

IgG
Research areas

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
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 second

Blocking/Dilution Buffer: 5% NFDM/TBST
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 seconds

Blocking/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).
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.
Anti-LAG-3 antibody [EPR20294-77] (ab209238)

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