Anti-Monoacylglycerol Lipase/MGL antibody [RM1358]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver (PMID : 30034939).

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-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Human adipose tissue tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human adipose tissue.

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

• IP

Lab

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Monoacylglycerol Lipase/MGL was immunoprecipitated from 0.35 mg DU145 whole cell lysate with ab325000 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325000 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : DU145 (human prostate carcinoma epithelial cell) whole cell lysate
Lane 2 : ab325000 IP in DU145 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325000 in DU145 whole cell lysate

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

DU145 (human prostate carcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab325000 at 1/30 IP in DU145 whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325000 in DU145 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

Observed band size: 30 kDa

false

Exposure time: 10s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat 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-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Rat adipose tissue tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat adipose tissue.

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-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse 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-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Immunohistochemical analysis of paraffin-embedded Mouse adipose tissue tissue labeling Monoacylglycerol Lipase/MGL with ab325000 at 1/2000 (0.251 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse adipose tissue.

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

• IP

Lab

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Monoacylglycerol Lipase/MGL was immunoprecipitated from 0.35 mg Rat brown fat whole tissue lysate with ab325000 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325000 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat brown fat whole tissue lysate
Lane 2 : ab325000 IP in Rat brown fat whole tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325000 in rat brown fat whole tissue lysate

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

Exposure time : 111 seconds.

All lanes:

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

Rat brown fat whole tissue lysate at 10 µg

Lane 2:

ab325000 at 1/30 IP in Rat brown fat whole tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325000 in rat brown fat whole tissue 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

Observed band size: 30 kDa

false

Exposure time: 111s

• IP

Lab

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

Monoacylglycerol Lipase/MGL was immunoprecipitated from 0.35 mg Mouse brown fat whole tissue lysate with ab325000 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325000 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brown fat whole tissue lysate
Lane 2 : ab325000 IP in Mouse brown fat whole tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325000 in mouse brown fat whole tissue lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

Mouse brown fat whole tissue lysate at 10 µg

Lane 2:

ab325000 at 1/30 IP in Mouse brown fat whole tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325000 in mouse brown fat whole tissue 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

Observed band size: 30 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

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

The expression of MGL is down-regulated in response to JZL treatment (PMID : 21802006).

Low expression : HCT-116 (PMID : 22349814).

The recombinant monoclonal antibody (ab325000, left) at 1 : 1000 dilution exhibited greater sensitivity compared to the polyclonal antibody (ab24701, right) at the same dilution.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002)(1/5000)(124KDa).

Exposure time : Left : 37 seconds Right : 180 seconds (This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range)

All lanes:

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

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

Lane 2:

DU145 treated with 1 uM JZL184 for 4 hours whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HCT 116 (human colorectal carcinoma 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: 33 kDa,124 kDa

true

• WB

Lab

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

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

All lanes:

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

Human breast tissue lysate at 20 µg

Lane 2:

Human fat tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Mouse brown fat tissue lysate at 20 µg

Lane 5:

Mouse liver tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Lane 7:

Rat brown fat tissue lysate at 20 µg

Lane 8:

Rat liver tissue lysate at 20 µg

Lane 9:

Rat heart 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: 33 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

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

The expression of MGL is down-regulated in response to JZL treatment (PMID : 21802006).

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

Exposure time : Lane 1-2 : 26 seconds Lane 3-5 : 70 seconds

All lanes:

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

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

Lane 2:

DU145 treated with 1 uM JZL184 for 4 hours whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

LNCaP (human prostate carcinoma 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: 33 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (AB325000)

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

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

All lanes:

Western blot - Anti-Monoacylglycerol Lipase/MGL antibody [RM1358] (ab325000) at 1/1000 dilution

Lane 1:

B16-F10 (mouse skin melanoma cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

B16-F10 transfected with siRNA specifically targeting MGL 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 1/20000 dilution

Observed band size: 33 kDa,36 kDa

false

Exposure time: 180s