Product nameAnti-RAP1 antibody [4c8/1]
See all RAP1 primary antibodies
DescriptionMouse monoclonal [4c8/1] to RAP1
Tested applicationsSuitable for: IP, WB, ELISA, Flow Cyt, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Human Rap1 protein.
- In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; HepG2; A431.
Storage instructionsShipped 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 bufferConstituent: PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab14404 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.|
|ELISA||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab91366 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration.|
FunctionActs both as a regulator of telomere function and as a transcription regulator. Involved in the regulation of telomere length and protection as a component of the shelterin complex (telosome). In contrast to other components of the shelterin complex, it is dispensible for telomere capping and does not participate in the protection of telomeres against non-homologous end-joining (NHEJ)-mediated repair. Instead, it is required to negatively regulate telomere recombination and is essential for repressing homology-directed repair (HDR), which can affect telomere length. Does not bind DNA directly: recruited to telomeric double-stranded 5'-TTAGGG-3' repeats via its interaction with TERF2. Independently of its function in telomeres, also acts as a transcription regulator: recruited to extratelomeric 5'-TTAGGG-3' sites via its association with TERF2 or other factors, and regulates gene expression. When cytoplasmic, associates with the I-kappa-B-kinase (IKK) complex and acts as a regulator of the NF-kappa-B signaling by promoting IKK-mediated phosphorylation of RELA/p65, leading to activate expression of NF-kappa-B target genes.
Tissue specificityUbiquitous. Highly expressed.
Sequence similaritiesBelongs to the RAP1 family.
Contains 1 BRCT domain.
Contains 1 Myb-like domain.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus. Cytoplasm. Chromosome. Chromosome > telomere. Associates with chromosomes, both at telomeres and in extratelomeric sites. Also exists as a cytoplasmic form, where it associates with the IKK complex.
- Information by UniProt
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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.
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
ab14404 staining RAP1 in the 184A1 epithelial cell line from Human mammary glands by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol/acetone (1:1), permeabilized with 1% Triton X-100 in PBS and blocked with 1% BSA for 10 minutes at 21°C. Samples were incubated with primary antibody (1/50 in PBS + 1% BSA) for 1 hour at 21°C. An undiluted Alexa Fluor®488-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody.
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.