Anti-LRBA/BGL antibody [EPR29133-75]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-LRBA/BGL antibody [EPR29133-75] (AB322661)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling LRBA/BGL with ab322661 at 1/50 (9.85 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 cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : A375
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-LRBA/BGL antibody [EPR29133-75] (AB322661)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A375 (human malignant melanoma epithelial cell, Left) / HeLa(human cervical adenocarcinoma epithelial cell, Right) cells labelling LRBA/BGL with ab322661 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : A375.

• WB

Supplier Data

Western blot - Anti-LRBA/BGL antibody [EPR29133-75] (AB322661)

"

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

Low expression : A375

Samples are non-boiled as boiling may cause protein aggregation.

The identity of the bands lower than 319 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lanes1-2 : 26 seconds, lanes3-4 : 10 seconds

"

All lanes:

Western blot - Anti-LRBA/BGL antibody [EPR29133-75] (ab322661) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 30 µg

Lane 2:

HeLa transfected with siRNA specifically targeting LRBA whole cell lysate at 30 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 40 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: 319 kDa,330 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Anti-LRBA/BGL antibody [EPR29133-75] (AB322661)

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

The expression of LRBA is upregulated in response to LPS treatment (PMID : 11254716).

Samples are non-boiled as boiling may cause protein aggregation.

The identity of the bands lower than 319 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-LRBA/BGL antibody [EPR29133-75] (ab322661) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 treated with 1ug/ml LPS for 24 hours 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: 319 kDa,330 kDa,124 kDa

false

Exposure time: 26s