重组Anti-LAG-3抗体[EPR20294-77]

• ICC/IF

Supplier Data

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

• IP

Supplier Data

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.

All lanes:

Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

Predicted band size: 57 kDa

Observed band size: 54 kDa,70 kDa

false

• WB

Supplier Data

Western blot - Anti-LAG-3 antibody [EPR20294-77] (AB209238)

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).

All lanes:

Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/1000 dilution

Lane 1:

Untreated mouse splenocytes whole cell lysate at 10 µg

Lane 2:

Mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 57 kDa

Observed band size: 54 kDa,70 kDa

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-LAG-3 antibody [EPR20294-77] (AB209238)

Blocking/Dilution Buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/10000 dilution

All lanes:

Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus, 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 57 kDa

Observed band size: 100 kDa

true

Exposure time: 1s