Anti-MBOAT7 antibody [EPR30427-503]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Human bladder cancer tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on human bladder cancer.

The primary antibody 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Panel A : HEK-293T (human embryonic kidney epithelial cell) transfected with a MBOAT7 expression vector containing a his tag Panel B : HEK-293T (human embryonic kidney epithelial cell) transfected with a SPZ1 expression vector containing a his tag Panel C : HEK-293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a MBOAT7 expression vector containing a his tag, no staining on (B) HEK-293T transfected with a SPZ1 expression vector containing a his tag and (C) HEK-293T cells transfected with empty vector containing a myc-His-tag.

The primary antibody 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on human testis.

The primary antibody 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Panel A : NIH : OVCAR-3 (human ovary adenocarcinoma epithelial cell) cell pellet Panel B : PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet Panel C : SW620 (human colorectal adenocarcinoma epithelial cell) cell pellet Panel D : THP-1 (human monocytic leukemia monocyte) cell pellet tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) NIH : OVCAR-3 and (B) PC-3, no staining in (C) SW620 and (D) THP-1.

The primary antibody 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on human breast cancer.

The primary antibody 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HT-1197(human urinary bladder carcinoma cell);THP-1(human monocytic leukemia monocyte) cells labelling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HT-1197 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : THP-1. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-KDEL mouse monoclonal antibody was used to counterstain tubulin at 1/1000 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 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : Nearly no staining in human liver.

The primary antibody 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling LENG4 with ab325494 at 1/500 (1.066 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : Nearly no staining in human kidney.

The primary antibody 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

• WB

Lab

Western blot - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

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

Low expression : lymph node, liver

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MBOAT7 antibody [EPR30427-503] (ab325494) at 1/1000 dilution

Lane 1:

Human adrenal gland tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Lane 3:

Human lymph node tissue lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 40 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-MBOAT7 antibody [EPR30427-503] (AB325494)

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

Low expression : THP-1, SW620

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MBOAT7 antibody [EPR30427-503] (ab325494) at 1/1000 dilution

Lane 1:

A-172 (human brain glioblastoma cell ) whole cell lysate at 40 µg

Lane 2:

HT-1197 (human urinary bladder carcinoma cell ) whole cell lysate at 40 µg

Lane 3:

NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 40 µg

Lane 6:

SW620 (human colorectal adenocarcinoma 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: 40 kDa,36 kDa

false

Exposure time: 180s