Anti-IFNGR1 antibody [EPR24127-89]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

Flow cytometric analysis of 4T1 (mouse mammary gland carcinoma epithelial cell, left)/ EL4 (mouse lymphoma T lymphocyte, right) cells labelling IFNGR1 with ab280353 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Low expression : 4T1(PMID : 12154047) Gated on viable cells.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 cells labelling IFNGR1 with ab280353 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in EL4 cell line. Low expression control : 4T1 (PMID : 12154047) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

Flow cytometric analysis of Mouse blood cells cells labelling IFNGR1 with ab280353 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• IP

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Immunoprecipitation - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

IFNGR1 was immunoprecipitated from 0.35 mg EL4 (mouse lymphoma T lymphocyte), whole cell lysate 10 ug with ab280353 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280353 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : EL4 (mouse lymphoma T lymphocyte), whole cell lysate 10 ug

Lane 2 : ab280353 IP in EL4 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab280353 in EL4 whole cell lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-IFNGR1 antibody [EPR24127-89] (ab280353)

Predicted band size: 54 kDa

Observed band size: 70-95 kDa

false

• WB

Lab

Western blot - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602).

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and IFNGR1-KO homozygous mice (Strain ID : T012667).

All lanes:

Western blot - Anti-IFNGR1 antibody [EPR24127-89] (ab280353) at 1/1000 dilution

Lane 1:

Untreated IFNGR1 KO mouse lung tissue whole lysate at 20 µg

Lane 2:

IFNGR1 KO mouse lung tissue treated with 1μg/mL for 16 hours whole lysate at 20 µg

Lane 3:

Mouse lung tissue treated with 1μg/mL for 16 hours whole lysate at 20 µg

Lane 4:

Untreated IFNGR1 KO mouse spleen tissue whole lysate at 20 µg

Lane 5:

IFNGR1 KO mouse spleen tissue treated with 1μg/mL for 16 hours whole lysate at 20 µg

Lane 6:

Mouse spleen tissue treated with 1μg/mL for 16 hours whole lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 80 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IFNGR1 antibody [EPR24127-89] (AB280353)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Low expression : 4T1(PMID : 12154047)

Exposure time : 74 seconds

All lanes:

Western blot - Anti-IFNGR1 antibody [EPR24127-89] (ab280353) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 20 µg

Lane 2:

4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 54 kDa

Observed band size: 70-95 kDa

false