Anti-Butyrylcholinesterase antibody [EPR26630-54]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human liver. The section was incubated with ab317016 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded (A) LNCaP (human prostate carcinoma epithelial cell) cell pellet (A) PC3 (human prostate cancer cell line) cell pellet tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on LNCaP cell pellet (image A), no staining on PC3 cell pellet (image B). The section was incubated with ab317016 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on plasma of human tonsil. The section was incubated with ab317016 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression tissue : weak staining on human kidney. The section was incubated with ab317016 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on plasma of human placenta. The section was incubated with ab317016 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Clear cell carcinoma of human kidney tissue labeling Butyrylcholinesterase with ab317016 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on plasma of clear cell carcinoma of human kidney. The section was incubated with ab317016 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IP

Supplier Data

Immunoprecipitation - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Butyrylcholinesterase was immunoprecipitated from 0.35 mg DU 145 (human prostate carcinoma epithelial cell) whole cell lysate with ab317016 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317016 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : DU 145 (human prostate carcinoma epithelial cell) whole cell lysate
Lane 2 : ab317016 IP in DU 145 (human prostate carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317016 in DU 145 whole cell lysate

All lanes:

Immunoprecipitation - Anti-Butyrylcholinesterase antibody [EPR26630-54] (ab317016) at 1/30 dilution

All lanes:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate

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

false

Exposure time: 76s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Butyrylcholinesterase with ab317016 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on mouse kidney. The section was incubated with ab317016 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-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunofluorescent analysis of 100% Methanol fixed 0.1% Triton X-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell); NIH/3T3 (mouse embryonic fibroblast) cells labelling Butyrylcholinesterase with ab317016 at 1/50 (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 Hepa1-6 cell line and no staining in NIH/3T3 cell line. The counterstain was observered with magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Negative control : NIH/3T3.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Butyrylcholinesterase with ab317016 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver. The section was incubated with ab317016 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

Supplier Data

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Low expression : PC-3 (PMID : 29966864).

Negative control : NIH/3T3 (PMID : 7842168)

BCHE is a glycoprotein. The molecular weight observed is consistent with what have been described in literature (PMID : 16434405, 24618259).

The identity of the lower MW band at approximately 25 kDa in lane 5 is unknown.

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

All lanes:

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (ab317016) at 1/1000 dilution

Lane 1:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Hepa1-6 (mouse hepatoma 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 1/20000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Low expression : kidney (PMID : 25448037, 7886701).

Butyrylcholinesterase is a glycoprotein and the 160KD band represents the dimer(PMID : 22889087).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16434405, 24618259).

The identity of the lower MW bands at approximately 50 kDa are unknown.

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

All lanes:

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (ab317016) at 1/1000 dilution

Lane 1:

Human plasma at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human kidney 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 1/20000 dilution

Observed band size: 76 kDa,160 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

Low expression : kidney (PMID : 25448037, 7886701), testis.

Butyrylcholinesterase is a glycoprotein and the 160KD band represents the dimer(PMID : 22889087).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16434405, 24618259).

The identity of the lower MW band at approximately 37 kDa in lane 2 is unknown.

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

All lanes:

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (ab317016) at 1/1000 dilution

Lane 1:

Mouse serum at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

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 1/20000 dilution

Observed band size: 76 kDa,160 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (AB317016)

This antibody does not cross-react with human Acetylcholinesterase.

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

All lanes:

Western blot - Anti-Butyrylcholinesterase antibody [EPR26630-54] (ab317016) at 1/1000 dilution

Lane 1:

His-tagged human Butyrylcholinesterase recombinant protein at 10 ng

Lane 2:

His-tagged human Acetylcholinesterase protein 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: 90 kDa

false

Exposure time: 1s