Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 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 in human liver cancer (PMID : 23979840,26422581,27241665) is observed.

The section was incubated with ab254262 for 15 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 solution 2) 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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Negative control : No staining in human cerebrum (PMID : 23979840)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 at 1/1000 dilution (0.5 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

The section was incubated with ab254262 for 15 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 solution 2) 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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Negative control : No staining in human colon (PMID : 23979840).

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 at 1/1000 dilution (0.5 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

The section was incubated with ab254262 for 15 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 solution 2) 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).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell line) cells labelling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in HepG2 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counter stain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 at 1/1000 dilution (0.5 μg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human liver (PMID : 23979840, 26422581, 27241665) is observed.

The section was incubated with ab254262 for 15 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 solution 2) for 20 mins.

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

• Flow Cyt

Unknown

Flow Cytometry - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Flow cytometric analysis of HepG2 (Human hepatocellular carcinoma epithelial cell line) cells labelling Asialoglycoprotein Receptor 1/HL-1 antibody with ab254262 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• IP

Unknown

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Asialoglycoprotein Receptor 1/HL-1 antibody was immunoprecipitated from 0.35 mg HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10μg with ab254262 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254262 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10μg

Lane 2 : ab254262 IP in HepG2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254262 in HepG2 whole cell lysate.

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

Exposure time : 7 seconds.

The band at around 100kDa is described as dimers (PMID : 7663158).

All lanes:

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (ab254262)

Predicted band size: 33 kDa

Observed band size: 40-50 kDa

false

• WB

Unknown

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (AB254262)

Negative control : human brain.

The diffuse bands between 40-50 kDa are likely due to glycosylation (PMID : 30965276).

The expression profile is consistent with what has been described in the literature (PMID : 27241665, 29858079).

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

All lanes:

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22642-17] (ab254262) at 1/1000 dilution

Lane 1:

Human adult liver tissue lysate at 20 µg

Lane 2:

Human brain tissue lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Lane 3:

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

Predicted band size: 33 kDa

Observed band size: 40-50 kDa

false