ET1605-16

p53 Recombinant Rabbit Monoclonal Antibody [SY010-6]

cat.: ET1605-16

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality:	Monoclonal
Clone number:	SY010-6

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: 44 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within N-terminal human p53.
Positive control:	HT-29 cell lysate, A431 cell lysate, MDA-MB-231 cell lysate, HEK-293 cell lysate, MDA-MB-468 cell lysate, Hela, MCF-7, HepG2, human colon carcinoma tissue.
Subcellular location:	Cytoplasm, Nucleus, Endoplasmic reticulum, Mitochondrion matrix.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:1,000-1:5,000 1:50-1:200 1:50-1:200 1:50-1:200
Uniprot #:	SwissProt: P04637 Human
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 protein 53 Tumor protein p53 Tumor suppressor p53

Images

Fig1: Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (ET1605-16) at 1/5,000 dilution.

Lane 1: HT-29 cell lysate
Lane 2: A431 cell lysate
Lane 3: Saos-2 cell lysate (negative)

Lysates/proteins at 15 µg/Lane.

Predicted band size: 44 kDa
Observed band size: 53 kDa

Exposure time: 5 minutes;

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 (ET1605-16) 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.

Fig2: Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (ET1605-16) at 1/1,000 dilution.

Lane 1: MDA-MB-231 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: MDA-MB-468 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 44 kDa
Observed band size: 53 kDa

Exposure time: 1 minute; 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 (ET1605-16) 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.

	Fig3: ICC staining of p53 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: ICC staining of p53 in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: ICC staining of p53 in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-p53 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1605-16, 1/50) for 30 minutes 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”.