p53 Recombinant Rabbit Monoclonal Antibody [SA39-07]
cat.: ET1601-13
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, ChIP
Clonality: Monoclonal
Clone number: SA39-07
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: 53 kDa
Isotype: IgG
Immunogen: Recombinant protein within human p53 aa 1-100.
Positive control: 293T cell lysate, 293T.
Subcellular location: Cytoplasm, Nucleus, Endoplasmic reticulum, Mitochondrion matrix
Recommended Dilutions:
  WB
  IF-Cell
  ChIP

1:5,000
1:400
Use 0.5~2 μg for 25 μg of chromatin.
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
ET1601-13_1.jpg Fig1: Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (ET1601-13) at 1/5,000 dilution.

Lane 1: 293T cell lysate
Lane 2: Saos-2 cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 43 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 (ET1601-13) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1601-13_2.jpg Fig2: Immunocytochemistry analysis of 293T cells labeling p53 with Rabbit anti-p53 antibody (ET1601-13) at 1/400 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-p53 antibody (ET1601-13) at 1/400 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.
ET1601-13_3.jpg Fig3: Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 treated with UV for 40minutes then recover for 3 hours cells and either p53 (ET1601-13) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.