All lanes : Anti-GC1q R antibody [EPR23238-107] (ab270032) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 31 kDa

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 15 secs.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20100866).

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling GC1q R with ab270032 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing mitochondrial staining in HeLa cell line. ab33985 Anti-COX IV antibody (human) was used to counterstain tubulin at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue). 

Negative controls:
-ve control 1: ab270032 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab150077 at 1/1000 dilution followed by ab33985 at 1/1000 dilution.

 

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling GC1q R with ab270032 at 1/2000 (0.248 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebral cortex (PMID: 23924515). The section was incubated with ab270032 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GC1q R with ab270032 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

All lanes : Anti-GC1q R antibody [EPR23238-107] (ab270032) at 1/5000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : MCF7 ( human breast adenocarcinoma epithelial cell) whole cell
Lane 4 : Rat spleen lysate
Lane 5 : Mouse spleen lysate
Lane 6 : WI-38 (human fetal lung fibroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 31 kDa

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

Exposure time: Lanes 1-5: 26 secs; Lane 6: 10 secs.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20100866).

All lanes : Anti-GC1q R antibody [EPR23238-107] (ab270032) at 1/5000 dilution

Lane 1 : Human lung cancer lysate
Lane 2 : Human spleen lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 31 kDa

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

Exposure time: 26 secs.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20100866).

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labeling GC1q R with ab270032 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labeling GC1q R with ab270032 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing mitochondrial staining in C2C12 cell line. ab33985 Anti-COX IV antibody (human) was used to counterstain tubulin at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue). 

Negative controls:
-ve control 1: ab270032 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab150077 at 1/1000 dilution followed by ab33985 at 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling GC1q R with ab270032 at 1/2000 (0.248 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebral cortex (PMID: 9414106). The section was incubated with ab270032 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling GC1q R with ab270032 at 1/2000 (0.248 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebral cortex (PMID: 9414106). The section was incubated with ab270032 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).