Anti-Albumin antibody [RM1208]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Albumin antibody [RM1208] (AB320751)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Albumin with ab320751 at 1/100 (4.87 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in HepG2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : HeLa.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Albumin antibody [RM1208] (AB320751)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Albumin with ab320751 at 1/2000 (0.244 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma in human tonsil. The section was incubated with ab320751 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Albumin antibody [RM1208] (AB320751)

Immunohistochemical analysis of paraffin-embedded (A) Rat non-perfused fixed kidney (B) Rat perfused fixed kidney tissue labeling Albumin with ab320751 at 1/2000 (0.244 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma in (A) rat non-perfused fixed kidney, no staining on (B) rat perfused fixed kidney. The section was incubated with ab320751 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Albumin antibody [RM1208] (AB320751)

Immunohistochemical analysis of paraffin-embedded (A) Mouse non-perfused fixed kidney (B) Mouse perfused fixed kidney tissue labeling Albumin with ab320751 at 1/2000 (0.244 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma in (A) mouse non-perfused fixed kidney and (B) mouse perfused fixed kidney. The section was incubated with ab320751 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-Albumin antibody [RM1208] (AB320751)

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

Negative control : Hela (PMID : 10476216 and 8314088) and NIH/3T3 (PMID : 4335066).

In lane 2 and 4, FBS was removed from the media for few hours before sample preparation for WB as albumin from FBS can affect WB results.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Albumin antibody [RM1208] (ab320751) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa in FBS-free media for 48 hours whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 in FBS-free media for 16 hours 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 1/100000 dilution

Observed band size: 70 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Albumin antibody [RM1208] (AB320751)

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

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Albumin antibody [RM1208] (ab320751) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Hepa1-6 (mouse hepatoma 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 1/100000 dilution

Observed band size: 68 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Albumin antibody [RM1208] (AB320751)

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

Exposure time : Lane 1 : 6 seconds; Lanes 2-3 : 4 seconds

All lanes:

Western blot - Anti-Albumin antibody [RM1208] (ab320751) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Human plasma lysate at 2 µg

Lane 4:

Rat plasma lysate at 2 µg

Lane 5:

Mouse plasma lysate at 2 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 1/100000 dilution

Observed band size: 68 kDa

false

• WB

Supplier Data

Western blot - Anti-Albumin antibody [RM1208] (AB320751)

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

All lanes:

Western blot - Anti-Albumin antibody [RM1208] (ab320751) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Lane 7:

Mouse kidney tissue lysate at 20 µg

Lane 8:

Mouse spleen tissue lysate at 20 µg

Lane 9:

Rat brain tissue lysate at 20 µg

Lane 10:

Rat kidney tissue lysate at 20 µg

Lane 11:

Rat heart tissue lysate at 20 µg

Lane 12:

Rat spleen 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 1/100000 dilution

Observed band size: 68 kDa

false

Exposure time: 6s