Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

Immunofluorescent analysis of 100% Methanol-fixed HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Asialoglycoprotein Receptor 1/HL-1 with at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line is observed. Methanol fixation is preferred.

ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Asialoglycoprotein Receptor 1/HL-1 with ab254261 at 1/2000 dilution (0.25 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human liver (PMID : 23979840,26422581,27241665). Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling Asialoglycoprotein Receptor 1/HL-1 with ab254261 at 1/2000 dilution (0.25 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human liver cancer (PMID : 23979840,26422581,27241665).No staining in human colon (PMID : 23979840). Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

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

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Asialoglycoprotein Receptor 1/HL-1 with ab254261 at 1/2000 dilution (0.25 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• IP

Unknown

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

Asialoglycoprotein Receptor 1/HL-1 was immunoprecipitated from 0.35 mg HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with ab254261 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254261 1/1000 dilution (0.45 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10μg
Lane 2 : ab254261 IP in HepG2 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254261 in HepG2 whole cell lysate.

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

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (ab254261)

Predicted band size: 33 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

Asialoglycoprotein Receptor 1/HL-1 was immunoprecipitated from 0.35 mg human liver lysate with ab254261 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254261 at 1/1000 dilution (0.45 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human liver lysate 10μg.

Lane 2 : ab254261 IP in human liver lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254261 in human liver lysate.

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

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (ab254261)

Predicted band size: 33 kDa

false

• WB

Unknown

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

The diffuse bands between 40-50 kDa are likely due to glycosylation.

The expression profile is consistent with what has been described in the literature (PMID : 27241665, 29858079, 22898823).

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

Exposure times : Lane 1 : 5.5 seconds; Lane 2 : 15 seconds.

All lanes:

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (ab254261) at 1/1000 dilution

Lane 1:

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

Lane 2:

Human liver 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

Predicted band size: 33 kDa

Observed band size: 40-50 kDa

false

• WB

Unknown

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (AB254261)

The diffuse bands between 40-50 kDa are likely due to glycosylation.

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

This blot was developed using a higher sensitivity ECL substrate.

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

All lanes:

Western blot - Anti-Asialoglycoprotein Receptor 1/HL-1 antibody [EPR22639-68] (ab254261) at 1/1000 dilution

All lanes:

Huh7 (human hepatocellular 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 1/20000 dilution

Predicted band size: 33 kDa

Observed band size: 40-50 kDa

false

Exposure time: 3min