Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

Clone CAN-R9(IHC)-56-2 (ab216646) has been successfully conjugated by Abcam. This image was generated using Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (Alexa Fluor^® 647). Please refer to ab202639 for protocol details.

ab202639 staining Wilms Tumor Protein in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab202639 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

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] - BSA and Azide free (AB216646)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

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] - BSA and Azide free (AB216646)

Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Wilms Tumor Protein with ab89901 at 1/200 dilution (0.1μg)/ Red. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

Immunocytochemsitry/Immunofluorescence analysis of K562 cells labelling 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

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). 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

Immunocytochemsitry/Immunofluorescence analysis of K562 cells labelling Wilms Tumor Protein (green) with unpurified 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

Clone CAN-R9(IHC)-56-2 (ab216646) has been successfully conjugated by Abcam. This image was generated using Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (Alexa Fluor^® 488). Please refer to ab202635 for protocol details.

ab202635 staining Wilms Tumor Protein in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab202635 at 1/200 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HepG2 cells fixed with 100% methanol (5min).

• IHC-P

AbReview51230****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

Immunohistochemical analysis of fromaldehyde fixed, paraffin embedded rat testis tissue sections, labelling 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 4ºC. The tissues were blocked with 10% Serum for 30 minutes at 25ºC. The secondary used was a Donkey CY3^® conjugate at a 1/200 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

This image is courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab89901).

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (AB216646)

This data was developed using ab89901, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/wilms-tumor-protein-antibody-can-r9ihc-56-2-ab89901'>ab89901</a>) 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

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