WT1 Antibody (6F-H2) - BSA Free

Western Blot: WT1 Antibody (6F-H2) [NB110-60011] - Hypomethylation of the Intron 1 CpG island results in WT1 expression. RNA was isolated from K562 and U937 cells growing under atmospheric conditions (Normox) and from U937 cells growing in 1% O2 (Hypox) and was analyzed by RT-PCR using primers that span exon 5 of the WT1 mRNA. Ribosomal RNA 36B4 is used as a positive control, and molecular weight markers (MW) are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0119837), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: WT1 Antibody (6F-H2) [NB110-60011] - Staining of human kidney glomerulus tissue. Heat mediated antigen retrieval was performed by heating in citrate buffer (pH 6) at 95C for 20 minutes. Image from verified customer review.

Western Blot: WT1 Antibody (6F-H2) [NB110-60011] - Whole cell protein from human K562, HEK293 and kidney tissue was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-WT1 in block buffer and detected with an anti-mouse HRP secondary antibody using chemiluminescence.

Immunohistochemistry: WT1 Antibody (6F-H2) [NB110-60011] - Analysis of Wilms Tumor 1 in human renal cancer using DAB with hematoxylin counterstain.

Simple Western: WT1 Antibody (6F-H2) [NB110-60011] - Lane view shows a specific band for WT1 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: WT1 Antibody (6F-H2) - BSA Free [NB110-60011] - Hypomethylation of Intron 1 CpG island results in WT1 expression. (C) Cell lysates from K562, U937, & HL60 cells treated with CoCl2, 5-azacytidine, or untreated controls analyzed by WB using an antibody against WT1. Histone H3 used as a loading control. Size of molecular weight markers is in kDa. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0119837), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF, IHC, IP
• Clone: 6F-H2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

WT1 Antibody (6F-H2) - BSA Free Summary

• Immunogen: Human recombinant Wilms Tumor 1 protein (WT1) (residues 1-181). [Swiss-Prot: P19544]
• Localization: Nuclear, Cytoplasm
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: WT1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence reported in scientific literature (PMID 25576053)
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Frozen 1: 400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation 2-10 ug/ml lysate
  • Simple Western 1:50
  • Western Blot 2 ug/ml
• Application Notes: In Western blot, a doublet is observed at approx. 55 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. In immunostaining, this target may be found localized to nucleus/nucleolus and cytoplasm of cells (WT1 shuttles between nucleus and cytoplasm - PMID: 14681305). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Hek293 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 68 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 55 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  293 Whole Cell Lysate
  K-562 Whole Cell Lysate

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for WT1 Antibody (6F-H2) - BSA Free

• AWT1
• GUD
• NPHS4GUD
• WAGR
• Wilms tumor 1
• Wilms tumor protein
• WIT-2
• WT1
• WT33

Background

Wilms' tumor 1 (WT1) gene inactivation is responsible for almost 20% of Wilms' tumour/developmental renal tumour or nephroblastoma cases and besides playing important role in cellular development and survival, this homodimeric transcription factor has been shown to involve in certain pathologies also. WT1 belongs to EGR C2H2-type zinc-finger protein family and interacts with several proteins including WTIP, HNRNPU, U2AF2, CITED2, ZNF224, WTAP, SRY, AMER1, RBM4 and actively translating polysomes (detected in nuclear ribonucleoprotein particles) for exerting its activities. WT1 binds to DNA sequence 5'-CGCCCCCGC-3' and regulates the expression of several target genes including EPO. It plays essential role in urogenital system development and in carcinogenesis, it has been suggested as tumor suppressor as well as oncogenic protein. Defects in WT1 have been linked to Wilms tumor 1 (WT1), Frasier syndrome (FS), Denys-Drash syndrome (DDS), nephrotic syndrome type 4 (NPHS4), Meacham syndrome (MEACHS), mesothelioma malignant (MESOM).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for WT1 Antibody (NB110-60011)(16)

Publications using NB110-60011

• Miyazaki Y, Orisaka M, Fujita Y et al. Steroidogenic differentiation of human amniotic membrane-derived mesenchymal stem cells into a progesterone-/androgen-producing cell lineage by SF-1 and an estrogen-producing cell lineage by WT1−KTS Frontiers in Endocrinology 2024-09-18 [PMID: 39359415]
• Yang X, Chen Z, Luo Z et al. STING deletion alleviates podocyte injury through suppressing inflammation by targeting NLRP3 in diabetic kidney disease Cellular signalling 2023-06-15 [PMID: 37329999] (IHC, Mouse)
• Hu J, Zhang Z, Hu H et al. LRH-1 activation alleviates diabetes-induced podocyte injury by promoting GLS2-mediated glutaminolysis Cell proliferation 2023-04-13 [PMID: 37057309] (IHC, Mouse)
• Cao Y, Chen Z, Hu J et al. Mfn2 Regulates High Glucose-Induced MAMs Dysfunction and Apoptosis in Podocytes via PERK Pathway Frontiers in Cell and Developmental Biology 2021-12-20 [PMID: 34988075]
• Loubalova Z, Fulka H, Horvat F et al. Formation of spermatogonia and fertile oocytes in golden hamsters requires piRNAs Nature Cell Biology 2021-09-06 [PMID: 34489573]
• M Wang, L Xiong, LJ Jiang, YZ Lu, F Liu, LJ Song, F Xiang, XL He, F Yu, SY Shuai, WL Ma, H Ye miR-4739 mediates pleural fibrosis by targeting bone morphogenetic protein 7 EBioMedicine, 2019-03-05;41(0):670-682. 2019-03-05 [PMID: 30850350]
• Horne SJ, Vasquez J, Guo Y et al. Podocyte-Specific Loss of Kruppel-Like Factor 6 Increases Mitochondrial Injury in Diabetic Kidney Disease Diabetes Aug 16 2018 [PMID: 30115650] (IHC, Mouse)
• Fan Ying, Li Xuezhu, Xiao Wenzhen et al. BAMBI elimination enhances alternative TGF-b signaling and glomerular dysfunction in diabetic mice. Diabetes 2015 [PMID: 25576053] (ICC/IF, Mouse)
• Jeitany M, Pineda JR, Liu Q et al. A preclinical mouse model of glioma with an alternative mechanism of telomere maintenance (ALT). Int. J Cancer. 2014-08-30 [PMID: 25175359]
• McCarty G, Awad O, Loeb DM. WT1 directly regulates expression of vascular endothelial growth factor and is a mediator of tumor response to hypoxia. The Journal of Biological Chemistry. 2011-10-26 [PMID: 22030397]
• Rauscher III, FJ et al. Characterization of monoclonal antibodies directed to the amino-terminus of the WT1, Wilms' tumor suppressor protein. Hybridoma;17(2):191-8. 1998-04-01 [PMID: 9627060]
• Loubalova Z, Fulka H, Horvat F, et al. Golden hamster piRNAs are necessary for early embryonic development and establishment of spermatogonia bioRxiv 2021-01-28 (ICC/IF, Hamster)
• Pruett N, Singh A, Shankar A Et Al. Normal Mesothelial Cell Lines Newly Derived from Human Pleural Biopsy Explants Am. J. Physiol. Lung Cell Mol. Physiol. 2020-07-29 [PMID: 32726133] (Human)
• McCarty G, Loeb DM. Hypoxia-Sensitive Epigenetic Regulation of an Antisense-Oriented lncRNA Controls WT1 Expression in Myeloid Leukemia Cells. PLoS ONE. 2015-03-21 [PMID: 25794157] (WB, Human)
  
  Details:
  WT1 antibody used for WB on total cell lysates from K562, U937 and HL60 that were treated with CoCl2, 5-azacytidine, or untreated controls (Figure 2C).WT1 antibody used at a dilution of 1:1,000 for WB on total cell lysates from K562, U937 and HL60 that were treated with CoCl2, 5-azacytidine, or untreated controls (Figure 2C).
• Wang CH Development of Class II-agnostic CAR T Cell Therapy Targeting WT1 Peptide Presented by Diverse HLA class II Molecules Thesis 2022-01-01
• Medina Rangel P, Cross E, Liu C et al. Cell Cycle and Senescence Regulation by Podocyte Histone Deacetylase 1 and 2 Journal of the American Society of Nephrology : JASN 2022-11-22 [PMID: 36414418]

Reviews for WT1 Antibody (NB110-60011) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Human and Mouse.

reviewed by: Verified Customer WB Human and Mouse 10/17/2019

Summary

• Application: Western Blot
• Sample Tested: 293T whole cell lysate,Liver
• Species: Human and Mouse
• Lot: B-4

reviewed by: Santhosh Sivajothi IHC-P Human 05/22/2018

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Human kidney tissue
• Species: Human
• Lot: A-5

Comments

• Comments: Heat mediated antigen retrieval was performed by heating in citrate buffer (pH6) at 95C for 20 minutes

reviewed by: Evren Azeloglu WB Human 04/28/2014

Summary

• Application: Western Blot
• Sample Tested: Immortalized podocytes
• Species: Human

FAQs for WT1 Antibody (NB110-60011). (Showing 1 - 2 of 2 FAQs).

1. I just wondering this antibody (WT1 antibody (6F-H2)[Alexa Flour488], when I use it to staining tissue do I need incubate in second antibody or not?
   • Our product NB110-60011AF488 is directly conjugated with an Alexa Fluor 488 dye, so you do not need to use any secondary antibody with this product to detect the antibody. The dye labeled to this antibody excites at 490nm and emits at 525nm.
2. In NB800-PC6, were the 293 cells transfected with WT1 expression plasmids? We have 293 cell lysates and it showed negative WT1 expression. Just want to make sure if our antibody is not working.
   • This protein lysate was derived from human HEK293 cells (without any transfection) and it is a whole cell lysate. WT1's mRNA is expressed significantly in HEK294 cells protein atlas site and given the fact that our Wilms Tumor 1 antibody (6F-H2) [NB110-60011] showed positive bands on HEK293 cells lysate, you should be good for considering this lysate as a positive control for WT1.

Bioinformatics

• Gene Symbol: WT1