Anti-Prosurfactant Protein C antibody [EPR21012-72] - BSA and Azide free (Capture)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody [EPR21012-72] - BSA and Azide free (Capture) (AB259667)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T cells transfected with an empty expression vector containing a myc-His-tag^®; HEK-293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag^®; HEK-293T cells transfected with a human SFTPC expression vector containing a myc-His-tag^® cells labeling Prosurfactant Protein C with ab259667 at 1/5000 dilution following ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in 293T cells (shown in green) transfected a mouse SFTPC expression vectorcontaining a myc-His-tag^® and 293T cells transfected a human SFTPC expression vector containing a myc-His-tag^®. 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). The nuclear counterstain was DAPI (Blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Prosurfactant Protein C antibody [EPR21012-72] - BSA and Azide free (Capture) (AB259667)

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 ab259667 at 1/10000 dilution (0.01 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 [EPR21012-72] - BSA and Azide free (Capture) (AB259667)

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 ab259667 at 1/10000 dilution (0.01 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.

• sELISA

Unknown

Sandwich ELISA - Anti-Prosurfactant Protein C antibody [EPR21012-72] - BSA and Azide free (Capture) (AB259667)

Representative standard curve from corresponding SimpleStep ELISA® Kit (ab252366).

• WB

Lab

Western blot - Anti-Prosurfactant Protein C antibody [EPR21012-72] - BSA and Azide free (Capture) (AB259667)

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 [EPR21012-72] - BSA and Azide free (Capture) (ab259667) 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

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Exposure time: 4s