HA722703

Phospho-p53 (S33) Recombinant Rabbit Monoclonal Antibody [JE41-97]

cat.: HA722703

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE41-97

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 53 kDa
Isotype:	IgG
Immunogen:	Synthetic phospho-peptide corresponding to residues surrounding Ser33 of Human p53 aa 1-50 / 393.
Positive control:	HT-29 cell lysate, HT-29 treated with 100ng/mL Nocodazole for 18 hours cell lysate, HT-29 cells treated with 100ng/mL Nocodazole for 18 hours, mouse liver tissue, human breast cancer tissue, human colon cancer tissue, human colon tissue, rat liver tissue.
Subcellular location:	Cytoplasm, Nucleus, PML body, Endoplasmic reticulum, Mitochondrion matrix, cytoskeleton, microtubule organizing center, centrosome.
Recommended Dilutions: WB IF-Cell IHC-P	1:1,000 1:100 1:200
Uniprot #:	SwissProt: P04637 Human \| P02340 Mouse \| P10361 Rat
Alternative names:	Antigen NY-CO-13 BCC7 Cellular tumor antigen p53 FLJ92943 LFS1 Mutant tumor protein 53 p53 p53 tumor suppressor P53_HUMAN Phosphoprotein p53 Tp53 Transformation related protein 53 TRP53 tumor antigen p55 Tumor protein 53 Tumor protein p53 Tumor suppressor p53

Images

Fig1: Western blot analysis of Phospho-p53 (S33) on different lysates with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) at 1/1,000 dilution.

Lane 1: HT-29 cell lysate
Lane 2: HT-29 treated with 100ng/mL Nocodazole for 18 hours cell lysate
Lane 3: HT-29 treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 15 µg/Lane.

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 3 minutes; 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 (HA722703) 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 HT-29 cells treated with 100ng/mL Nocodazole for 18 hours labeling Phospho-p53 (S33) with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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-Phospho-p53 (S33) antibody (HA722703) 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA722703) at 1/200 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 cancer tissue with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA722703) at 1/200 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 colon cancer tissue with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA722703) at 1/200 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 human colon tissue with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA722703) at 1/200 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 liver tissue with Rabbit anti-Phospho-p53 (S33) antibody (HA722703) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA722703) at 1/200 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.

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