Immunofluorescence analysis of WT1 was performed using 70% confluent log phase SW-480 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with WT1 Rabbit Polyclonal Antibody (PA516879) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from the C-terminus of human Wilm's tumor protein|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1-3 µg/ml|
* 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.
|Immunohistochemistry (IHC)||See 4 publications below|
PA5-16879 targets Wilms Tumor Protein in IHC (P) applications and shows reactivity with Human, mouse, and Rat samples.
The PA5-16879 immunogen is a synthetic peptide derived from the C-terminus of human Wilm's tumor protein.
Wilm's Tumor (WT), a sporadic and familial childhood kidney tumor, is genetically heterogeneous. Wilm's tumor is associated with mutations of WT1, a zinc-finger transcription factor that is essential for the development of the metanephric kidney and the urogenital system. The WT1 gene is normally expressed infetal kidney and mesothelioma, and its expression has been suggested as a marker for Wilm's tumor and mesothelioma.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Diagnostic pitfalls of differentiating desmoplastic small round cell tumor (DSRCT) from Wilms tumor (WT): overlapping morphologic and immunohistochemical features.
PA5-16879 was used in immunohistochemistry to study the difficulty in differentially diagnosing desmoplastic small round cell tumor and Wilms tumor
|Arnold MA,Schoenfield L,Limketkai BN,Arnold CA||The American journal of surgical pathology (38:1220)||2014|
Intrauterine growth restriction leads to a dysregulation of Wilms' tumour supressor gene 1 (WT1) and to early podocyte alterations.
PA5-16879 was used in immunohistochemistry to study the dysregulation of the Wilm's tumor suppressor gene-1 in response to uterine growth restriction and the significance for renal disease susceptibility
|Menendez-Castro C,Hilgers KF,Amann K,Daniel C,Cordasic N,Wachtveitl R,Fahlbusch F,Plank C,Dötsch J,Rascher W,Hartner A||Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association (28:1407)||2013|
Proliferation and maturation of microvessels in arteriovenous malformations--expression patterns of angiogenic and cell cycle-dependent factors.
PA5-16879 was used in immunohistochemistry to study the patterns of proliferation and maturation of microvessels in arteriovenous malformations
|Meijer-Jorna LB,van der Loos CM,Teeling P,de Boer OJ,Florquin S,van der Horst CM,van der Wal AC||Journal of cutaneous pathology (39:610)||2012|
Connexin 43 regulates epicardial cell polarity and migration in coronary vascular development.
PA5-16879 was used in immunohistochemistry to study the role of Connexin 43 in epicardial cell polarity and migration during coronary vascular development
|Rhee DY,Zhao XQ,Francis RJ,Huang GY,Mably JD,Lo CW||Development (Cambridge, England) (136:3185)||2009|