重组Anti-Prosurfactant蛋白C抗体[EPR19839-8]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized transfected HEK-293T cells labelling Prosurfactant Protein C with ab270521 at 1/5000 (0.1166μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used as a counterstain at 1/100 (5μg/ml). Confocal image showing cytoplasmic staining in 293T cells (shown in green) transfected 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®. 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).

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®

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Flow cytometry (intracellular) analysis of transfected 293T cells. Cells were fixed with 4% PFA for 10 minutes and permeabilised with 90% methanol before staining for Prosurfactant Protein C in 293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag® using antibody ab270521 at 1/5000 dilution (0.01 ug) (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. Cells were co-stained with anti-myc conjugated to Alexa Fluor®647.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Prosurfactant Protein C with ab270521 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The Pulmonary surfactant-associated protein C is a peptide secreted by the alveolar type II pneumocytes of the lung (PMID : 16769199). Negative control : No staining on mouse spleen. The section was incubated with ab270521 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung tissue labeling Prosurfactant Protein C with ab270521 at 1/100 dilution followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining on mouse lung is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Prosurfactant Protein C with ab270521 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Type II alveolar cells in mouse lung. The section was incubated with ab270521 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Prosurfactant Protein C was immunoprecipitated from 0.35 mg Mouse lung tissue lysate 1with ab270521 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab270521 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse lung tissue lysate 10 ug.

Lane 2 : ab270521 IP in Mouse lung tissue lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270521 in mouse lung tissue lysate.

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

Exposure time : 6 seconds.

All lanes:

Immunoprecipitation - Anti-Prosurfactant Protein C antibody [EPR19839-8] (ab270521)

Predicted band size: 21 kDa,67 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

Blocking and diluting buffer and concentration : This target has a precursor which contains an exceedingly hybrophobic region of 34 amino acids that comprises most of the mature Spftc, and a unique poly-valine domain. The staining pattern of our antibody is similar with published paper (PMID : 20378731 and PMID : 22461427). 15 seconds Exposure time :

All lanes:

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839-8] (ab270521) at 1/1000 dilution

All lanes:

Mouse lung tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-Prosurfactant Protein C antibody [EPR19839-8] (AB270521)

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 [EPR19839-8] (ab270521) 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='/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