Anti-beta Galactosidase antibody [EPR29591-746]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a Beta-galactosidase expression vector containing a his tag and (B) HEK-293T transfected with empty vector containing a his tag labeling beta Galactosidase with ab324616 at 1/1000 (0.476 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) HEK-293T transfected with a Beta-galactosidase expression vector containing a his tag and no signal in (B) HEK-293T transfected with empty vector containing a his tag.
The section was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling beta Galactosidase with ab324616 at 1/1000 (0.476 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing positive staining in RAW 264.7 cells phagocytized Escherichia coli (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).
A : RAW 264.7 cells
B : Raw 264.7 cells phagocytized Escherichia coli and treated with 0.5% Glucose for 18h
C : Raw 264.7 cells phagocytized Escherichia coli

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

Immunohistochemical analysis of paraffin-embedded (A) Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellets; (B) Raw 264.7 cells phagocytized Escherichia coli and treated with 0.5% Glucose for 18h; (C) Raw 264.7 cells phagocytized Escherichia coli cell pellets labeling beta Galactosidase with ab324616 at 1/1000 (0.476 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (C) Raw 264.7 cells phagocytized Escherichia coli cell pellets, and no signal in (A) Raw 264.7 cell pellets and (B) Raw 264.7 cells phagocytized Escherichia coli and treated with 0.5% Glucose for 18h.
The section was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

beta Galactosidase was immunoprecipitated from 0.35 mg E. coli lysate with ab324616 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324616 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-beta Galactosidase antibody [EPR29591-746] (ab324616) at 1/1000 dilution

Lane 1:

E. coli lysate (Input) at 10 µg

Lane 2:

ab324616 at 1/30 IP in E. coli lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 116 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (ab324616) at 1/1000 dilution

Lane 1:

His-tagged Beta-galactosidase recombinant fragment at 10 ng

Lane 2:

His-tagged Regulatory protein GAL4 recombinant fragment at 10 ng

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: 30 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

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

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

All lanes:

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (ab324616) at 1/5000 dilution

Lane 1:

Untreated E.coli lysate at 4 µg

Lane 2:

E.coli treated with 1mM IPTG for 3 hours lysate at 4 µg

Lane 3:

E.coli treated with 0.5% Glucose for 18 hours lysate at 4 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: 116 kDa,36 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (AB324616)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at a 1/5000 dilution; Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) at a 1/100000 dilution.

All lanes:

Western blot - Anti-beta Galactosidase antibody [EPR29591-746] (ab324616) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Beta-galactosidase expression vector containing a His-tag, 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 1/100000 dilution

Observed band size: 116 kDa,15 kDa

false

Exposure time: 3s