Anti-RAP1 antibody [4c8/1] (ab14404)
Key features and details
- Mouse monoclonal [4c8/1] to RAP1
- Suitable for: IP, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG2b
Overview
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Product name
Anti-RAP1 antibody [4c8/1]
See all RAP1 primary antibodies -
Description
Mouse monoclonal [4c8/1] to RAP1 -
Host species
Mouse -
Tested applications
Suitable for: IP, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Human Rap1 protein.
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Positive control
- In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; HepG2; A431.
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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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
4c8/1 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG2b -
Research areas
Images
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All lanes : Anti-RAP1 antibody [4c8/1] (ab14404) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
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RAP1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to RAP1 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, Hela whole cell extract 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab14404.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 55kDa; RAP1 -
Overlay histogram showing HeLa cells stained with ab14404 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab14404, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.