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Western Blot (WB) (Western blot analysis of WT1 expression in K562 cell lysate (MBS179182).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-WT1 monoclonal antibody overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for WT1 )

Rabbit anti-Human, Mouse Wilms Tumor Protein Monoclonal Antibody | anti-WT1 antibody

Anti-Wilms Tumor Protein Rabbit Monoclonal Antibody

Gene Names
WT1; GUD; AWT1; WAGR; WT33; NPHS4; WIT-2
Reactivity
Human, Mouse
Applications
Flow Cytometry, Functional Assay, Immunofluorescence, Immunohistochemistry, Immunocytochemistry, Western Blot
Purity
Affinity-chromatography
Synonyms
Wilms Tumor Protein; Monoclonal Antibody; Anti-Wilms Tumor Protein Rabbit Monoclonal Antibody; Wilms tumor protein; WT33; WT1; anti-WT1 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse
Clonality
Monoclonal
Purity/Purification
Affinity-chromatography
Form/Format
Liquid
Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Sequence Length
288
Applicable Applications for anti-WT1 antibody
Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB)
Application Notes
WB: 1:500-1:1000
IHC: 1:50-1:200
ICC/IF: 1:50-1:200
FC/FACS: 1:50
Immunogen
A synthesized peptide derived from human Wilms Tumor Protein
Subcellular Localization
Nucleus. Nucleus, nucleolus. Cytoplasm. Shuttles between nucleus and cytoplasm.
Tissue Specificity
Expressed in the kidney and a subset of hematopoietic cells.
Preparation and Storage
Store at -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Western blot analysis of WT1 expression in K562 cell lysate (MBS179182).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-WT1 monoclonal antibody overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for WT1 )

Western Blot (WB) (Western blot analysis of WT1 expression in K562 cell lysate (MBS179182).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-WT1 monoclonal antibody overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for WT1 )

Immunohistochemistry (IHC)

(Immunohistochemical analysis of paraffin-embedded mouse kidney, using Wilms Tumor Protein Antibody(MBS179182)WT1 was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-WT1 Antibody (MBS179182)overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Immunohistochemical analysis of paraffin-embedded mouse kidney, using Wilms Tumor Protein Antibody(MBS179182)WT1 was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-WT1 Antibody (MBS179182)overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Related Product Information for anti-WT1 antibody
Transcription factor that plays an important role in cellular development and cell survival. Regulates the expression of numerous target genes, including EPO. Plays an essential role for development of the urogenital system. Recognizes and binds to the DNA sequence 5'-CGCCCCCGC-3'. It has a tumor suppressor as well as an oncogenic role in tumor formation. Function may be isoform-specific: isoforms lacking the KTS motif may act as transcription factors. Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing. Isoform 1 has lower affinity for DNA, and can bind RNA.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
49188 MW
NCBI Official Full Name
Wilms tumor protein isoform A
NCBI Official Synonym Full Names
Wilms tumor 1
NCBI Official Symbol
WT1
NCBI Official Synonym Symbols
GUD; AWT1; WAGR; WT33; NPHS4; WIT-2
NCBI Protein Information
Wilms tumor protein
UniProt Protein Name
Wilms tumor protein
Protein Family
UniProt Gene Name
WT1

NCBI Description

This gene encodes a transcription factor that contains four zinc-finger motifs at the C-terminus and a proline/glutamine-rich DNA-binding domain at the N-terminus. It has an essential role in the normal development of the urogenital system, and it is mutated in a small subset of patients with Wilms tumor. This gene exhibits complex tissue-specific and polymorphic imprinting pattern, with biallelic, and monoallelic expression from the maternal and paternal alleles in different tissues. Multiple transcript variants have been described. In several variants, there is evidence for the use of a non-AUG (CUG) translation initiation codon upstream of, and in-frame with the first AUG. Authors of PMID:7926762 also provide evidence that WT1 mRNA undergoes RNA editing in human and rat, and that this process is tissue-restricted and developmentally regulated. [provided by RefSeq, Mar 2015]

Uniprot Description

Transcription factor that plays an important role in cellular development and cell survival (PubMed:7862533). Recognizes and binds to the DNA sequence 5'-GCG(T/G)GGGCG-3' (PubMed:7862533, PubMed:17716689, PubMed:25258363). Regulates the expression of numerous target genes, including EPO. Plays an essential role for development of the urogenital system. It has a tumor suppressor as well as an oncogenic role in tumor formation. Function may be isoform-specific: isoforms lacking the KTS motif may act as transcription factors (PubMed:15520190). Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing (PubMed:16934801). Isoform 1 has lower affinity for DNA, and can bind RNA (PubMed:19123921).

Research Articles on WT1

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Product Notes

The WT1 wt1 (Catalog #AAA179182) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Wilms Tumor Protein Rabbit Monoclonal Antibody reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's Wilms Tumor Protein can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB). WB: 1:500-1:1000 IHC: 1:50-1:200 ICC/IF: 1:50-1:200 FC/FACS: 1:50. Researchers should empirically determine the suitability of the WT1 wt1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Wilms Tumor Protein, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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