Anti-Metabotropic Glutamate Receptor 7/MGLUR7 抗体 [EPR24530-11] (ab302530)

All lanes : Anti-Metabotropic Glutamate Receptor 7/MGLUR7 antibody [EPR24530-11] (ab302530) at 1/1000 dilution

Lane 1 : Mouse hippocampus tissue lysate
Lane 2 : Mouse spinal cord tissue lysate
Lane 3 : Mouse heart tissue lysate
Lane 4 : Mouse skeletal muscle tissue lysate
Lane 5 : Mouse spleen tissue lysate
Lane 6 : Rat hippocampus tissue lysate
Lane 7 : Rat heart tissue lysate
Lane 8 : Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 102 kDa
Observed band size: 260 kDa why is the actual band size different from the predicted?


Exposure time: 81 seconds

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

Negative controls: heart, skeletal muscle, spleen (PMID: 12052533).

260-kDa GRM7 dimer is observed. The molecular weight is consistent with literature (PMID: 33767338, 19109906).

Anti-Metabotropic Glutamate Receptor 7/MGLUR7 antibody [EPR24530-11] (ab302530) at 1/1000 dilution + SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 102 kDa
Observed band size: 260 kDa why is the actual band size different from the predicted?


Exposure time: 92 seconds

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

260-kDa GRM7 dimer is observed. The molecular weight is consistent with literature (PMID: 33767338, 19109906).

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Metabotropic Glutamate Receptor 7/MGLUR7 with ab302530 at 1/1000 dilution (0.5 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse hippocampus (PMID: 11007882, PMID: 9295396). The section was incubated with ab302530 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling Metabotropic Glutamate Receptor 7/MGLUR7 with ab302530 at 1/1000 dilution (0.5 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat hippocampus (PMID: 11007882, PMID: 9295396). The section was incubated with ab302530 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) labeling Metabotropic Glutamate Receptor 7 / MGLUR7 with ab302530 at 1/500 dilution (1 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on (A) HEK-293T transfected with a Myc-His tagged GRM7 construct, no staining on (B) HEK-293T transfected with empty plasmid. The section was incubated with ab302530 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labeling Metabotropic Glutamate Receptor 7/MGLUR7 with ab302530 at 1/1000 dilution (0.5 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on mouse cardiac muscle. The section was incubated with ab302530 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling Metabotropic Glutamate Receptor 7/MGLUR7 with ab302530 at 1/1000 dilution (0.5 µg/mL) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on rat cardiac muscle. The section was incubated with ab302530 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat hippocampus (fresh) tissue labeling Metabotropic Glutamate Receptor 7 / MGLUR7 with AB302530 at 1/50 dilution (10.02 µg/mL) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Positive staining on rat hippocampus is observed. The nuclear counterstain was DAPI (Blue).

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

I

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat heart (fresh) tissue labeling Metabotropic Glutamate Receptor 7/MGLUR7 with AB302530 at 1/50 dilution (10.02 µg/mL)) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control: no staining on rat heart is observed (PMID 12052533). The nuclear counterstain was DAPI (Blue).

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

Metabotropic Glutamate Receptor 7/MGLUR7 was immunoprecipitated from 0.35 mg mouse hippocampus tissue lysate with AB302530 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB302530 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse hippocampus tissue lysate 10 µg (Inset)

Lane 2: AB302530 IP in Mouse hippocampus tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of AB302530 in mouse hippocampus tissue lysate

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

Exposure time: 3 minutes. 

Observed MW (kDa) 260,110. 

260-kDa GRM7 dimer and 110-kDa GRM7 monomer are observed (PMID: 33767338).

Metabotropic Glutamate Receptor 7/MGLUR7 was immunoprecipitated from 0.35 mg rat hippocampus tissue lysate with AB302530 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB302530 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Rat hippocampus tissue lysate 10 µg (Inset)

Lane 2: AB302530 IP in rat hippocampus tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of AB302530 in rat hippocampus tissue lysate

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

Exposure time: 3 minutes

Observed MW (kDa): 260, 110. 

260-kDa GRM7 dimer and 110-kDa GRM7 monomer are observed (PMID: 33767338).