Anti-Prosurfactant Protein C antibody [EPR19839]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Prosurfactant Protein C with ab211326 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control : no staining on mouse spleen. [PMID : 15186480]. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Flow cytometry (intracellular) analysis of transfected 293T cells. Cells were fixed with 4% PFA for 10 minutes and permeabilised with 0.1% Tween-20 before staining for Prosurfactant Protein C in 293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag® (Middle panel). Staining was compared with 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right panel) and with an isotype control, Rabbit monoclonal IgG (ab172730) (left panel). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody at a 1/5000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized transfected HEK-293T cells labelling Prosurfactant Protein C with ab211326 at 1/5000 dilution (0.207 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing cytoplasmic staining in 293T cells (shown in green) transfected with a mouse SFTPC expression vector containing a myc-His-tag®, showing no staining in 293T cells transfected a human SFTPC expression vector containing a myc-His-tag®. ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) at 1/100 (5μg/ml) was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Panel A : HEK-293T cells transfected with an empty expression vector containing a myc-His-tag®

Panel B : HEK-293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag®

Panel C : HEK-293T cells transfected with a human SFTPC expression vector containing a myc-His-tag®

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Prosurfactant Protein C with ab211326 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on alveolar type II cells of mouse lung is observed [PMID : 15186480].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung tissue labeling Prosurfactant Protein C with ab211326 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Cytoplasmic staining on alveolar type II cells of mouse lung is observed [PMID : 23967208].

The nuclear counterstain is DAPI (blue).

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.

• IP

Unknown

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

SP-C was immunoprecipitated from 0.35 mg of mouse lung lysate with ab211326 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab211326 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : Mouse lung lysate, 10ug (Input).

Lane 2 : ab211326 IP in mouse lung lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab211326 in mouse lung lysate.

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

Exposure time : 5 seconds.

The two bands detected correspond to 22 kda and 21 kda isoforms.

All lanes:

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR19839] (ab211326)

Predicted band size: 21 kDa

Observed band size: 21 kDa,22 kDa

false

• WB

Supplier Data

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

Blocking/Dilution buffer : 5% NFDM/TBST.

Negative control : mouse spleen. [PMID : 15186480].

All lanes:

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839] (ab211326) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 20 µg

Lane 2:

Mouse spleen lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa,89 kDa

Observed band size: 100 kDa,21 kDa,22 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839] (AB211326)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution (36KDa).

In Western blot, Anti-6X His tag® antibody [AD1.1.10] total protein control (1/1000) (30KDa)

All lanes:

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839] (ab211326) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human SFTPC expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a mouse SFTPC expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 21 kDa

false

Exposure time: 4s