Anti-Prosurfactant Protein C antibody (ab90716)
Key features and details
- Rabbit polyclonal to Prosurfactant Protein C
- Suitable for: ICC/IF, IHC-P, WB, IP
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Prosurfactant Protein C antibody
See all Prosurfactant Protein C primary antibodies -
Description
Rabbit polyclonal to Prosurfactant Protein C -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanIP MouseWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Normal human lung. ICC: A549 cells. IP: Mouse Lung tissue. WB: Mouse and human Lung Tissue Lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prosurfactant Protein C antibody (ab90716)
IHC image of Prosurfactant Protein C staining in a section of formalin-fixed paraffin-embedded normal human lung performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab90716, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-Prosurfactant Protein C antibody (ab90716)All lanes : Anti-Prosurfactant Protein C antibody (ab90716) at 1 µg/ml
Lane 1 : Lung (Human) Tissue Lysate
Lane 2 : Lung (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 25,28 kDa why is the actual band size different from the predicted?
Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
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Western blot - Anti-Prosurfactant Protein C antibody (ab90716)All lanes : Anti-Prosurfactant Protein C antibody (ab90716) at 1 µg/ml
Lane 1 : Lung (Human) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 25,28 kDa why is the actual band size different from the predicted?
Additional bands at: 45 kDa (possible non-specific binding)
Exposure time: 2 minutes
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Immunocytochemistry/ Immunofluorescence - Anti-Prosurfactant Protein C antibody (ab90716)ab90716 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab90716 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunoprecipitation - Anti-Prosurfactant Protein C antibody (ab90716)Prosurfactant Protein C was immunoprecipitated using 0.5mg Mouse Lung tissue, 5µg of Rabbit polyclonal to Prosurfactant Protein C and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Lung tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab90716.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 23kDa; Prosurfactant Protein C
