WT1 Antibody (6F-H2)

Images

 
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 ...read more
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. ...read more
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.

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Clone
6F-H2
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml

WT1 Antibody (6F-H2) 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
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 2 ug/ml
  • Simple Western 1:50
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Frozen 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation 2-10 ug/ml lysate
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. Separated by Size-Wes, Sally Sue/Peggy Sue. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID: 25576053).
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
WT1 Overexpression Lysate
Reviewed Applications
Read 2 Reviews rated 4
using
NB110-60011 in the following applications:

Publications
Read Publications using
NB110-60011 in the following applications:

  • 1 publication
  • IHC
    1 publication
  • WB
    1 publication

Packaging, Storage & Formulations

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

Alternate Names for WT1 Antibody (6F-H2)

  • 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.

Customers Who Viewed This Item Also Viewed...

NBP1-52823
Species: Hu, Rt, Po, Bv, Ca, Eq, GP, Rb, Sh
Applications: WB, ICC/IF, IHC, IHC-P
NB200-103
Species: Hu, Mu, Rt, Ye, Xp(-)
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready, Flow-IC
AF2818
Species: Hu
Applications: WB, ICC
NBP2-16389
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P, IP
H00005076-M01
Species: Hu, Mu
Applications: WB, ELISA, ICC/IF, IHC-P
NBP2-42210
Species: Hu
Applications: WB, Flow, IHC, IHC-P
PP-H7431-00
Species: Hu
Applications: WB
NBP2-37463
Species: Hu
Applications: WB, ELISA, Flow, IHC, IHC-P, CyTOF-ready
AF3075
Species: Hu
Applications: WB, Simple Western, ICC
NB110-61646
Species: Hu, Mu
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
NBP2-75624
Species: Hu, Mu, Rt
Applications: WB, Flow, IHC, IHC-P
NB300-223
Species: Hu, Mu, Rt, Po, Bv, Ca, Ch, Eq
Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P
AF3844
Species: Hu, Mu
Applications: IHC
NB600-1071
Species: Mu, Rt
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
NBP2-45545
Species: Hu
Applications: WB, IHC
NB300-750
Species: Hu, Mu, Rt, Fi, Mk, Pm, Sh
Applications: WB, ICC/IF, IHC, IP, ICC
AF5414
Species: Hu
Applications: WB, Simple Western
NB110-60011
Species: Hu, Mu
Applications: WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P, IP

Publications for WT1 Antibody (NB110-60011)(6)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 3 applications: ICC/IF, IHC, WB.


Filter By Application
ICC/IF
(1)
IHC
(1)
WB
(1)
All Applications
Filter By Species
Human
(1)
Mouse
(2)
All Species
Showing Publications 1 - 6 of 6.
Publications using NB110-60011 Applications Species
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) IHC Mouse
McCarty G, Loeb DM. Hypoxia-Sensitive Epigenetic Regulation of an Antisense-Oriented lncRNA Controls WT1 Expression in Myeloid Leukemia Cells. PLoS ONE. 2015 Mar 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).
WB Human
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) 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 Aug 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 Oct 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 Apr. [PMID: 9627060]

Reviews for WT1 Antibody (NB110-60011) (2) 42

Average Rating: 4
(Based on 2 reviews)
We have 2 reviews tested in 1 species: Human.

Reviews using NB110-60011:
Filter by Applications
IHC-P
(1)
WB
(1)
All Applications
Filter by Species
Human
(2)
All Species
Images Ratings Applications Species Date Details
Immunohistochemistry-Paraffin WT1 NB110-60011
Enlarge
reviewed by:
Santhosh Sivajothi
IHC-P Human 05/22/2018
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedHuman kidney tissue
SpeciesHuman
LotA-5

Comments

CommentsHeat mediated antigen retrieval was performed by heating in citrate buffer (pH6) at 95C for 20 minutes
Western Blot WT1 NB110-60011
Enlarge
reviewed by:
Evren Azeloglu
WB Human 04/28/2014
View

Summary

ApplicationWestern Blot
Sample TestedImmortalized podocytes
SpeciesHuman

Blocking

Blocking DetailsRockland casein blocking buffer, 2 hrs, RT

Primary Anitbody

Dilution Ratio1:1000 in 5% milk in TBS-T, overnight, 4ºC

Secondary Antibody

Secondary DescriptionThermo goat anti-rabbit HRP
Secondary Concentration1:2000 in 5% milk in TBS-T, 30min, RT

Details

Detection NotesThermo ECL, 2min

Product General Protocols

View specific protocols for WT1 Antibody (NB110-60011): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

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.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB110-60011AF350
Alexa Fluor 405 NB110-60011AF405
Alexa Fluor 488 NB110-60011AF488
Alexa Fluor 532 NB110-60011AF532
Alexa Fluor 594 NB110-60011AF594
Alexa Fluor 647 NB110-60011AF647
Alexa Fluor 700 NB110-60011AF700
Alexa Fluor 750 NB110-60011AF750
Biotin NB110-60011B
DyLight 350 NB110-60011UV
DyLight 405 NB110-60011V
DyLight 488 NB110-60011G
DyLight 550 NB110-60011R
DyLight 594 NB110-60011DL594
DyLight 650 NB110-60011C
DyLight 680 NB110-60011FR
DyLight 755 NB110-60011IR
FITC NB110-60011F
HRP NB110-60011H
Janelia Fluor 549 NB110-60011JF549
Janelia Fluor 646 NB110-60011JF646

Additional WT1 Products

Bioinformatics Tool for WT1 Antibody (NB110-60011)

Discover related pathways, diseases and genes to WT1 Antibody (NB110-60011). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for WT1 Antibody (NB110-60011)

Discover more about diseases related to WT1 Antibody (NB110-60011).
 

Pathways for WT1 Antibody (NB110-60011)

View related products by pathway.

PTMs for WT1 Antibody (NB110-60011)

Learn more about PTMs related to WT1 Antibody (NB110-60011).
 

Research Areas for WT1 Antibody (NB110-60011)

Find related products by research area.

Blogs on WT1.

Routine WT1 Antibody Screen Uncovers an Exciting New Cancer-Cleaning Enzyme
Enzyme antibodies are widely used in both apoptosis and cancer studies, as disruption of the proteins regulating apoptosis is known to lead to formation of tumor cells in certain cancers. We at Novus Biologicals are continually growing our enzyme anti...  Read full blog post.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Recent Reviews

4
5
0
4
2
3
0
2
0
1
0

Santhosh Sivajothi
05/22/2018
Application: IHC-P
Species: Human

Evren Azeloglu
04/28/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol WT1