* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Antigen affinity chromatography
0.05% sodium azide
-20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
PA1-029 detects nucleophosmin (NPM) from human, rat, and mouse samples.
PA1-029 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~ 38 kDa protein corresponding to human, mouse, and rat nucleophosmin in various cell lysates.
The PA1-029 immunogen is a synthetic peptide corresponding to amino acid residues 23-38 and 226-240 from human NPM.
Nucleophosmin (NPM) also called B23, nutramin and NO38 is a ubiquitously expressed phosphoprotein involved in ribosome assembly/transport, cytoplasmic/nuclear trafficking, regulation of DNA polymerase alpha activity, centrosome duplication, and regulation of p53. NPM continuously shuttles between the nucleus and cytoplasm. It has been shown to bind nucleic acid, prevent protein aggregation via its chaperon activities, protect enzymes during thermal denaturation, and facilitate renaturation of chemically denatured proteins. In its cellular structure role, there is evidence suggesting NPM is associated with the centrosome. It is the substrate of CDK2/cyclin E during duplication of centrosomes (cellular division).
Due to the NPM gene interaction with several tumor-associated chromosome translocations, NPM is thought to be a portion of several fusion proteins: NPM-ALK, NPM-RAR, and NPM-MLF1. While it is not thought to be part of the transforming potential of these fusion proteins, it is believed to act as the interface for oligomerization and oncogenic conversion of these tumor promoting fusion proteins.
Nucleophosmin Antibody (PA1-029) in WB PA1-029_WB.jpg
Western Blot with anti-Nucleophosmin Polyclonal Antibody (PA1-029)
Western blot analysis of Nucleophosmin (Pierce Product # PA1-029) was performed by loading 25ug of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a rabbit polyclonal antibody recognizing Nucleophosmin at a dilution of 1:500 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody (Pierce Product # 31460) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Pico (Product #34077).
Nucleophosmin Antibody (PA1-029) in IHC PA1-029_Immunohistochemistry_Human_Kidney-tissue.jpg
Immunohistochemistry with anti-Nucleophosmin Polyclonal Antibody (PA1-029)
Immunohistochemistry was performed on normal deparaffinized human Kidney tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:500 with a rabbit polyclonal antibody recognizing Nucleophosmin (Product #PA1-029) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)