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Synthetic peptide: AGSVEQTPKKPGLRRRQT, corresponding to C terminal amino acids 181-198 of Human p27 KIP 1.
Our Abpromise guarantee covers the use of ab7961 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent dilution.|
|IHC-P||1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use at an assay dependent dilution. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).Can be blocked with p27 KIP 1 peptide (ab8016).|
|Sandwich ELISA||Use a concentration of 0.1 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal to p27 KIP 1 (ab54563). For sandwich ELISA, use this antibody as Detection at 0.1µg/ml with Ab54563 as Capture.|
|ICC/IF||Use a concentration of 1 µg/ml.|
ICC/IF image of ab7961 stained Hek293 cells (ab7902). 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 (ab7961, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.
ab7961 staining 927 KIP1 in human tonsillar tissue sections by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde. Antigen retrieval was by heat mediation in citrate buffer (20 minutes at 100°C). Samples were incubated with primary antibody 1/25 (background reducer diluent) for 20 minutes at 25°C. An undiluted HRP polymer Goat anti mouse/rabbit IgG (ab6721) was used as secondary antibody.
Immunohistochemistical detection of p27 KIP 1 antibody (ab7961) on formaldehyde-fixed paraffin-embedded mouse cerebellum sections. Antigen retrieval step: Heat mediated. Buffer Used: Citric acid pH6.3. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody: 1/250 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti rabbit IgG Conjugation: Biotin. An excellent immunostaining pattern that is most evident in nuclei. However, a significant number of nuclei are negative for this Ab and the significance of this is as yet unknown.
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