Anti-Prosurfactant Protein C antibody [EPR28174-18]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human lung tissue staining Prosurfactant Protein C with ab312850 at a 1 : 2000 (0.26 ug/ml) dilution; Uteroglobin with ab307666 at 1 : 2000 dilution and MARCO with ab314646 at 1 : 100 (5.1 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Prosurfactant Protein C (magenta; Opal™690), anti-Uteroglobin (green; Opal™520) and anti-MARCO (gray; Opal™570) on human lung.
Panel B : anti-Prosurfactant Protein C staining pneumocytes in human lung.
Panel C : anti-Uteroglobin staining club cells in human lung.
Panel D : anti-MARCO staining alveolar macrophages in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab312850, ab307666 and ab314646 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval 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-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung. The section was incubated with ab312850 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-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human spleen. The section was incubated with ab312850 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

• IP

Supplier Data

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Prosurfactant Protein C was immunoprecipitated from 0.35 mg Human lung tissue lysate with ab312850 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab312850 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human lung tissue lysate
Lane 2 : ab312850 IP in Human lung tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab312850 in human lung tissue lysate.

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

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR28174-18] (ab312850) at 1/1000 dilution

All lanes:

Human lung tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 1s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat spleen. The section was incubated with ab312850 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-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse spleen. The section was incubated with ab312850 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-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse lung (PMID : 23967208). The section was incubated with ab312850 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-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Prosurfactant Protein C with ab312850 at 1/2000 (0.26 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat lung. The section was incubated with ab312850 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

• WB

Supplier Data

Western blot - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : cerebellum, liver, kidney, spleen. The expression profile/ molecular weight observed is consistent with what has been described in the reference (PMID : 27770432, PMID : 34469722). In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution. Exposure time : Lane 1 : 3 seconds, lanes 2-5 : 180 seconds

All lanes:

Western blot - Anti-Prosurfactant Protein C antibody [EPR28174-18] (ab312850) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human kidney tissue lysate at 20 µg

Lane 5:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 21 kDa

false

Exposure time: 180s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a Myc tag; 293T transfected with a human SFTPC containing a Myc tag cells labeling Prosurfactant Protein C with ab312850 at 1/500 dilution following ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing membrane and cytoplasmic staining in 293T cells transfected with a human SFTPC containing a Myc tag (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor^® 594) was used to counterstain at 1/100 dilution (Magenta).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag^® (Middle); 293T cells transfected with an empty expression vector containing a myc-His-tag^® (Right) cells labelling Prosurfactant Protein C with ab312850 at 1/50000 dilution (0.001 ug) / Middle and right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti-myc conjugated to Alexa Fluor^®647.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a human SFTPC expression vector containing a myc-His-tag^® (Middle); 293T cells transfected with an empty expression vector containing a myc-His-tag^® (Right) cells labelling Prosurfactant Protein C with ab312850 at 1/50000 dilution (0.001 ug) / Middle and right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti-myc conjugated to Alexa Fluor^®647.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a Myc tag; 293T transfected with a mouse SFTPC containing a Myc tag cells labeling Prosurfactant Protein C with ab312850 at 1/500 dilution following ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing membrane and cytoplasmic staining in 293T cells transfected with a mouse SFTPC containing a Myc tag (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor^® 594) was used to counterstain at 1/100 dilution (Magenta).

• WB

Lab

Western blot - Anti-Prosurfactant Protein C antibody [EPR28174-18] (AB312850)

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

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

All lanes:

Western blot - Anti-Prosurfactant Protein C antibody [EPR28174-18] (ab312850) 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='/en-us/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