WT1 Antibody (6F-H2) - BSA Free

Novus Biologicals | Catalog # NB110-60011

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Hamster

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 6F-H2

Format

BSA Free
View all WT1 Primary Antibodies »

Product Specifications

Immunogen

Human recombinant Wilms Tumor 1 protein (WT1) (residues 1-181). [Swiss-Prot: P19544]

Localization

Nuclear, Cytoplasm

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

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.

Description

Novus Biologicals Mouse WT1 Antibody (6F-H2) - BSA Free (NB110-60011) is a monoclonal antibody validated for use in IHC, WB, ICC/IF, Simple Western and IP. Anti-WT1 Antibody: Cited in 14 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for WT1 Antibody (6F-H2) - BSA Free

Western Blot: WT1 Antibody (6F-H2)BSA Free [NB110-60011]

Western Blot: WT1 Antibody (6F-H2)BSA Free [NB110-60011]

WT1-Antibody-6F-H2-Western-Blot-NB110-60011-img0007.jpg
Immunohistochemistry-Paraffin: WT1 Antibody (6F-H2) - BSA Free [NB110-60011]

Immunohistochemistry-Paraffin: WT1 Antibody (6F-H2) - BSA Free [NB110-60011]

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)BSA Free [NB110-60011]

Western Blot: WT1 Antibody (6F-H2)BSA Free [NB110-60011]

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) - BSA Free [NB110-60011]

Immunohistochemistry: WT1 Antibody (6F-H2) - BSA Free [NB110-60011]

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)BSA Free [NB110-60011]

Simple Western: WT1 Antibody (6F-H2)BSA Free [NB110-60011]

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

Western Blot: WT1 Antibody (6F-H2) - BSA Free [NB110-60011] -

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 (https://dx.plos.org/10.1371/journal.pone.0119837), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for WT1 Antibody (6F-H2) - BSA Free

Application
Recommended Usage

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.

Reviewed Applications

Read 3 reviews rated 4.3 using NB110-60011 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: WT1

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).

Long Name

Wilms Tumor 1

Alternate Names

GUD, WAGR, WIT-2, WT33

Gene Symbol

WT1

Additional WT1 Products

Product Documents for WT1 Antibody (6F-H2) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for WT1 Antibody (6F-H2) - BSA Free

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.

Citations for WT1 Antibody (6F-H2) - BSA Free

Customer Reviews for WT1 Antibody (6F-H2) - BSA Free (3)

4.3 out of 5
3 Customer Ratings
5 Stars
33%
4 Stars
67%
3 Stars
0%
2 Stars
0%
1 Stars
0%

Have you used WT1 Antibody (6F-H2) - BSA Free?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Customer Images

Showing  1 - 3 的 3 reviews Showing All
Filter By:
Clear Filters X
  • WT1 Antibody (6F-H2)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Liver and 293T whole cell lysate
    Species: Mouse and Human
    Verified Customer | Posted 10/17/2019
    WT1 expression in Mouse Liver.
    WT1 Antibody (6F-H2) - BSA Free NB110-60011
  • WT1 Antibody (6F-H2)
    Name: Santhosh Sivajothi
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Human kidney tissue
    Species: Human
    Verified Customer | Posted 05/22/2018
    Staining in kidney glomerulus.
    Heat mediated antigen retrieval was performed by heating in citrate buffer (pH6) at 95C for 20 minutes
    WT1 Antibody (6F-H2) - BSA Free NB110-60011
  • Name: Evren Azeloglu
    Application: Western Blot
    Sample Tested: Immortalized podocytes
    Species: Human
    Verified Customer | Posted 04/28/2014
    Western blot, immortalized human podocytes (10ug); 1:1000, overnight 4C
    WT1 Antibody (6F-H2) - BSA Free NB110-60011

There are no reviews that match your criteria.

Showing  1 - 3 的 3 reviews Showing All

Protocols

View specific protocols for WT1 Antibody (6F-H2) - BSA Free (NB110-60011):

WT1 Antibody (6F-H2):
Western Blot Protocol
1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 28 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS, 1 hour at room temperature.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the mouse anti-Wilm's Tumor 1 primary antibody (NB 110-60011) in blocking buffer and incubate 2 hours at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted mouse-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers
instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce, ECL).

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for WT1 Antibody (6F-H2) - BSA Free

Showing  1 - 2 的 2 FAQs Showing All

    • Q: 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?
      A: 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.

    • Q: 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.
      A: 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.
Showing  1 - 2 的 2 FAQs Showing All
View all FAQs for Antibodies