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.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues 23-38 and 226-240 of human NPM.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500 - 1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.