Anti-LGALS3BP antibody [EPR26644-269]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-LGALS3BP antibody [EPR26644-269] (AB322977)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling LGALS3BP with ab322977 at 1/100 (4.88 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 : IMR-32.
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-LGALS3BP antibody [EPR26644-269] (AB322977)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized IMR-32(human neuroblastoma neuroblast, Left) / HeLa(human cervical adenocarcinoma epithelial cell, Right) cells labelling LGALS3BP with ab322977 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 : IMR-32

• WB

Supplier Data

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (AB322977)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 33076448, PMID : 26136428).

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

All lanes:

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (ab322977) at 1/1000 dilution

Lane 1:

Human salivary gland tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human stomach tissue lysate at 20 µg

Lane 4:

Human brainstem tissue lysate at 20 µg

Lane 5:

Human small intestine tissue lysate at 20 µg

Lane 6:

Human colon 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: 37-100 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (AB322977)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 33076448, PMID : 26136428).

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

All lanes:

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (ab322977) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting LGALS3BP 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: 37-100 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (AB322977)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 33076448, PMID : 26136428).

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

All lanes:

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (ab322977) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

HeLa transfected with siRNA specifically targeting LGALS3BP 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: 75-100 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (AB322977)

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

Low expression : IMR-32, U-937, RKO.

LGALS3BP is a glycoprotein of approximately 37-100 kDa and detected as a 30-60 kDa band after treated with Peptide : N-glycosidase F (PNGase F).

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 33076448, PMID : 26136428).

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

All lanes:

Western blot - Anti-LGALS3BP antibody [EPR26644-269] (ab322977) at 1/1000 dilution

Lane 1:

Untreated HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 whole cell lysate treated with Peptide: N-glycosidase F (PNGase F) at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 7:

RKO (human colon 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 1/100000 dilution

Observed band size: 30-60 kDa,37-100 kDa,36 kDa

false

Exposure time: 15s