Anti-GBP2 antibody [EPR29773-529]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GBP2 antibody [EPR29773-529] (AB324736)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling GBP2 with ab324736 at 1/50 (9.98 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in RAW 264.7 cell line treated with IFN gamma (10 ng/ml) for 24 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

Supplier Data

Western blot - Anti-GBP2 antibody [EPR29773-529] (AB324736)

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

Low expression : testis

The samples are unboiled.

The identity of the higher MW band at approximately 250 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GBP2 antibody [EPR29773-529] (ab324736) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse testis tissue 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 1/100000 dilution

Observed band size: 66 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-GBP2 antibody [EPR29773-529] (AB324736)

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

The expression of GBP2 is upregulated in response to IFN gamma treatment (PMID : 35383115).

The expression of GBP2 is upregulated in response to LPS treatment (PMID : 24739961).

The samples are unboiled.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GBP2 antibody [EPR29773-529] (ab324736) at 1/2000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 2 µg

Lane 2:

NIH/3T3 treated with 100ng/ml IFN gamma for 48 hours whole cell lysate at 2 µg

Lane 3:

Untreated J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate at 2 µg

Lane 4:

J774A.1 treated with 200ng/ml IFN gamma and 50 ng/ml LPS for 24 hours whole cell lysate at 2 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 1/100000 dilution

Observed band size: 66 kDa,36 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-GBP2 antibody [EPR29773-529] (AB324736)

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

The expression of GBP2 is upregulated in response to IFN gamma treatment (PMID : 35383115).

The samples are unboiled.

The identity of the higher MW band at approximately 250 kDa and lower MW band at approximately 30 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GBP2 antibody [EPR29773-529] (ab324736) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg

Lane 2:

RAW 264.7 treated with 10ng/ml IFN gamma for 24 hours whole cell lysate at 10 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 1/100000 dilution

Observed band size: 66 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-GBP2 antibody [EPR29773-529] (AB324736)

Dot blot analysis of GBP2 using ab324736 at 1 : 1000 (0.499 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : LIF/His-tagged Mouse GBP2 recombinant fragment
Lane2 : LIF-tagged Mouse GBP1 recombinant fragment
Lane3 : LIF/His-tagged Mouse GBP3 recombinant fragment
Lane4 : His-tagged Mouse GBP4 recombinant fragment
Lane5 : His-tagged Mouse GBP5 recombinant fragment
Lane6 : LIF/His-tagged Mouse GBP6 recombinant fragment
Lane7 : His-tagged Mouse GBP7 recombinant fragment

Exposure time : 180 seconds.

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

This antibody does not cross-react with mouse GBP1, GBP3, GBP4, GBP5, GBP6 or GBP7 by dot blot.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution. In Dot Blot, Anti-LIF antibody [EPR21713-25] - BSA and Azide free staining at Capture dilution.

All lanes:

Dot Blot - Anti-GBP2 antibody [EPR29773-529] (ab324736) at 1/1000 dilution

Lane 1:

LIF/His-tagged Mouse GBP2 recombinant fragment

Lane 2:

LIF-tagged Mouse GBP1 recombinant fragment

Lane 3:

LIF/His-tagged Mouse GBP3 recombinant fragment

Lane 4:

His-tagged Mouse GBP4 recombinant fragment

Lane 5:

His-tagged Mouse GBP5 recombinant fragment

Lane 6:

LIF/His-tagged Mouse GBP6 recombinant fragment

Lane 7:

His-tagged Mouse GBP7 recombinant fragment

Secondary

All lanes:

Dot 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 1/100000 dilution

false

Exposure time: 180s