All lanes : Anti-Albumin antibody [EPR20195] (ab207327) at 1/2000 dilution

Lane 1 : Human liver lysate at 20 µg
Lane 2 : Mouse liver lysate at 20 µg
Lane 3 : Rat liver lysate at 20 µg
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

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

Predicted band size: 69 kDa
Observed band size: 69 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-3: 5 seconds; Lane 4/5: 1 second.

Negative control: HeLa (PMID: 10476216 and 8314088).

Lanes 1-4 : Anti-Albumin antibody [EPR20195] (ab207327) at 1/10000 dilution
Lanes 5-6 : Anti-Albumin antibody [EPR20195] (ab207327) at 1/2000 dilution

Lane 1 : Human plasma at 2 µg
Lane 2 : Rat plasma at 2 µg
Lane 3 : Mouse plasma at 2 µg
Lane 4 : Human serum at 2 µg
Lane 5 : Human fetal kidney lysate at 10 µg
Lane 6 : Human fetal spleen lysate at 10 µg

Secondary
Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 5-6 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 69 kDa
Observed band size: 69 kDa


Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-Albumin antibody [EPR20195] (ab207327) at 1/2000 dilution

Lane 1 : Mouse spleen lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat spleen lysate
Lane 4 : Rat kidney lysate
Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) 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: 69 kDa
Observed band size: 69 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-4: 1 second; Lane 5: 3 minutes; Lane 6: 5 seconds.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) or HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Albumin with ab207327 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on HepG2 cell line.

Negative control: HeLa (PMID: 10476216 and 8314088)).

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde-fixed permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Albumin with ab207327 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

Albumin was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab207327 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab207327 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HepG2 whole cell lysate, 10µg (Input).

Lane 2: ab207327 IP in HepG2 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207327 in HepG2 whole cell lysate.

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

Exposure time: 30 seconds.

Albumin antibody ab207327 was used with 3D Cell Culture Clearing Kit ab243299 to penetrate, stain and clear a 3D hepatocyte cell culture.

Blue: DAPI, Green: CD68, Yellow: Albumin, Red: Vimentin

Learn more about 3D cell culture and tissue clearing kits, reagents, and protocols designed to make it easier to stain 3D cell cultures and thick tissue sections and get more data from each valuable tissue section.