Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

ab89901 staining Wilms Tumor Protein in paraffin-embedded human ovarian serous adenocarcinoma tissue sections by Immunohistochemistry.

Antigen retrieval was by heat mediation using citrate buffer (pH 6.0) was performed. Samples were incubated with primary antibody at 1 : 500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on human ovarian serous adenocarcinoma (PMID : 11939727) is observed.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

ab89901 staining Wilms Tumor Protein in paraffin-embedded human kidney tissue sections by Immunohistochemistry.

Antigen retrieval was by heat mediation using citrate buffer (pH 6.0). Samples were incubated with primary antibody at 1 : 500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on human kidney glomerulus (PMID : 12898605) is observed.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Wilms tumor tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Wilms Tumor Protein with ab89901 at 1/200 dilution (0.1 μg)/ Red.

Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) / Black was used as the isotype control. Cells without incubation with primary antibody and secondary antibody / Blue was used as the unlabeled control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Immunocytochemsitry/Immunofluorescence analysis of K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling Wilms Tumor Protein (green) with purified ab89901 at 1/50.

Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

• IHC-P

AbReview51230****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Immunohistochemical analysis of fromaldehyde fixed, paraffin embedded rat testis tissue sections, labeling Wilms Tumor Protein using ab89901.

Heat mediated antigen retrival was performed using 10 mM sodium citrate and 0.05% Tween-20. Tissue sections were incubated with ab89901 at a 1/50 dilution for 12 hours at 4°C. The tissues were blocked with 10% serum for 30 minutes at 25°C. The secondary used was a Donkey CY3^® conjugate at a 1/200 dilution.

This image is courtesy of an anonymous Abreview.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Spermatogenic phenotypes of Rnf8;Scml2-dKO mice.

Immunostaining of testicular sections with WT1 (ab89901), a marker of Sertoli cells, and PLZF, a marker of undifferentiated spermatogonia.

For preparation of testicular paraffin blocks, testes of mutants and littermate controls were fixed with 4% paraformaldehyde at 4°C overnight. Testes were then dehydrated and embedded in paraffin. For histological analyses, 6 μm-thick paraffin sections were deparaffinized and autoclaved in Target Retrieval Solution at 121°C for 20 min. The sections were blocked with for 10 min at room temperature, and then incubated with primary antibodies at 4°C overnight.

Adams et al PLoS Genet. 2018 Feb 20;14(2):e1007233. doi: 10.1371/journal.pgen.1007233. eCollection 2018 Feb. Fig S1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Podocyte counting using WT1 staining.

Mouse kidney sections were stained with WT1 antibody ab89901 using immunohistochemistry. (A) Each glomeruli was identified and numbered using ImagePro premier imaging software. (B) A tracing was applied along the Bowman's capsule to assess the surface area of the glomerulus. (C) Each WT1 positive nucleus was identified and the nuclear size assessed.

Verma et al PLoS One. 2018 Jun 20;13(6):e0198013. doi: 10.1371/journal.pone.0198013. eCollection 2018. Fig S1. Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

ab89901 staining Wilms Tumor Protein in paraffin-embedded mouse testis tissue sections by Immunohistochemistry.

Antigen retrieval was by heat mediation using citrate buffer (pH 6.0). Samples were incubated with primary antibody at 1 : 500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on Sertoli cells in mouse testis (PMID : 21863216) is observed.

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Western blot : Rabbit Monoclonal [CAN-R9(IHC)-56-2] to Wilms Tumor Protein ab89901 staining at 1/500 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 50/61 kDa in Wild-type HAP1 cell lysates with no signal observed at this size in WT1 knockout HAP1 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/500 dilution

Lane 1:

Wild-type HAP1 at 20 µg

Lane 2:

WT1 knockout HAP1 at 20 µg

Lane 3:

K562 at 20 µg

Lane 4:

SH-SY5Y at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 61 kDa

false

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Blocking buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/100 dilution

All lanes:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 49 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Exposure time : Left image : 3 minutes
Right image : 15 seconds

This antibody shows low affinity in mouse sample.

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/500 dilution

Lane 1:

Human testis whole cell lysate. at 15 µg

Lane 2:

K562 (Human T cell leukemia T lymphocyte) whole cell lysates at 4 µg

Lane 3:

Mouse testis whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 49 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

WT1 expression varies in different cell lines.

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/5000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast). Whole cell lysates at 20 µg

Lane 2:

THP-1 (Human monocytic leukemia monocyte). Whole cell lysates at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma epithelial cell) Whole cell lysates at 20 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 49 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution

All lanes:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anto-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 49 kDa

Observed band size: 55 kDa

false

• WB

Unknown

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901)

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution

All lanes:

Ramos (Human Burkitt's lymphoma cell line) cell lysate

Predicted band size: 49 kDa

false