ET1610-45

Wilms Tumor Protein Recombinant Rabbit Monoclonal Antibody [SC06-41]

cat.: ET1610-45

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, mIHC
Clonality:	Monoclonal
Clone number:	SC06-41

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 49 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human WT1 aa 59-269 / 449.
Positive control:	K-562 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, K-562, mouse testis tissue, mouse kidney tissue, human fallopian tube tissue, human testis tissue, human ovary cancer tissue, rat testis tissue.
Subcellular location:	Nucleus, Cytoplasm, Nucleus speckle.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P mIHC	1:2,000-1:5,000 1:100-1:200 1:200 1:500-1:1,000 1:100
Uniprot #:	SwissProt: P19544 Human \| P22561 Mouse \| P49952 Rat
Alternative names:	WIT 2 WT 1 AWT1 FWT1 GUD NPHS4 WAGR Wilms tumor 1 Wilms Tumor Wilms tumor protein Wilms' tumor gene Wilms' tumor protein WIT2 WT WT1 WT1_HUMAN WT33

Images

Fig1: Western blot analysis of Wilms Tumor Protein on different lysates with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/2,000 dilution.

Lane 1: K-562 cell lysate
Lane 2: Mouse testis tissue lysate
Lane 2: Rat testis tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 49 kDa

Exposure time: 42 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-45) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Wilms Tumor Protein on different lysates with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/5,000 dilution.

Lane 1: K-562-si NT cell lysate
Lane 2: K-562-si Wilms Tumor Protein cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 50 kDa

Exposure time: 2 minutes 37 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-45) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of K-562 cells labeling Wilms Tumor Protein with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunofluorescence analysis of paraffin-embedded mouse testis tissue labeling Wilms Tumor Protein with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-45, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig5: Immunofluorescence analysis of paraffin-embedded mouse kidney tissue labeling Wilms Tumor Protein with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-45, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

	Fig6: Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-45) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-45) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-45) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig9: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-45) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig10: mIHC analysis of mouse kidney tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-Wilms Tumor Protein antibody (ET1610-45) at 1/100 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit (MH900206). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.