Immunohistochemical analysis of paraffin-embedded U266 cell pellets (left, high-expressing) and Daudi (right, low-expressing) cell pellets using p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
|SignalStain® Boost IHC Detection Reagent (HRP, Mouse) #8125||SignalStain® Boost IHC Detection Reagent (AP, Mouse) #31926|
|SignalStain® DAB Substrate Kit #8059||SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713|
|SignalStain® Vivid Purple Peroxidase Substrate Kit #96632|
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2010
revised June 2020
Protocol Id: 280
p27 Kip1 (SX53G8.5) Mouse mAb (IHC Formulated) recognizes endogenous levels of total p27 Kip1 protein.
Monoclonal antibody is produced by immunizing animals with recombinant human p27 Kip1.
p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin-2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).
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