ET1610-44

p63 Recombinant Rabbit Monoclonal Antibody [SC06-31]

cat.: ET1610-44

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	SC06-31

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: 77 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human p63 aa 1-250.
Positive control:	A431 cell lysate, MCF7 cell lysate, HaCaT cell lysate, Mouse skin tissue lysate, Rat skin tissue lysate, A431, human skin tissue, human breast tissue, human esophagus tissue, mouse esophagus tissue, rat esophagus tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:2,000 1:500-1:2,000 1:50-1:200 1:800-1:2,000 1:1,000
Uniprot #:	SwissProt: Q9H3D4 Human \| O88898 Mouse \| Q9JJP6 Rat
Alternative names:	AIS Amplified in squamous cell carcinoma B(p51A) B(p51B) Chronic ulcerative stomatitis protein CUSP DN p63 alpha 1 DNp63 EEC3 id:ibd3516 Keratinocyte transcription factor Keratinocyte transcription factor KET KET LMS MGC115972 MGC192897 NBP OFC8 OTTHUMP00000209732 OTTHUMP00000209733 OTTHUMP00000209734 OTTHUMP00000209735 OTTHUMP00000209737 OTTHUMP00000209738 OTTHUMP00000209739 OTTHUMP00000209740 OTTHUMP00000209741 OTTHUMP00000209742 OTTHUMP00000209743 OTTHUMP00000209744 p40 p51 P51/P63 p53-related protein p63 p53CP p63 P63_HUMAN p73H p73L RHS SHFM4 TAp63alpha TP53CP TP53L TP63 TP73L Transformation related protein 63 Transformation-related protein 63 Trp53rp1 Trp63 Tumor protein 63 Tumor protein p53-competing protein Tumor protein p53-like Tumor protein p63 Tumor protein p63 deltaN isoform delta Tumor protein p73 Tumor protein p73-like

Images

Fig1: Western blot analysis of p63 on different lysates with Rabbit anti-p63 antibody (ET1610-44) at 1/1,000 dilution.

Lane 1: A431 cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane) (low expression)
Lane 3: HaCaT cell lysate (15 µg/Lane)
Lane 4: Mouse skin tissue lysate (30 µg/Lane)
Lane 5: Rat skin tissue lysate (30 µg/Lane)

Predicted band size: 77 kDa
Observed band size: 60-70 kDa

Exposure time: 10 seconds; ECL: K1801;

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 (ET1610-44) at 1/1,000 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: Immunocytochemistry analysis of A431 (positive) and MCF7 (negative) labeling p63 with Rabbit anti-p63 antibody (ET1610-44) at 1/2,000 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-p63 antibody (ET1610-44) at 1/2,000 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-p63 antibody (ET1610-44) at 1/800 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 (ET1610-44) at 1/800 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-p63 antibody (ET1610-44) at 1/2,000 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 (ET1610-44) at 1/2,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-p63 antibody (ET1610-44) at 1/2,000 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 (ET1610-44) at 1/2,000 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-p63 antibody (ET1610-44) at 1/2,000 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 (ET1610-44) at 1/2,000 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-p63 antibody (ET1610-44) at 1/2,000 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 (ET1610-44) at 1/2,000 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: Flow cytometric analysis of A431 cells labeling p63. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-44, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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