p53 (TP53) (NM_000546) Human Over-expression Lysate

CAT#: LC400186

TP53 HEK293T cell transient overexpression lysate (as WB positive control)

Product Images

TP53 activity in the over-expression lysate (LY400186) was measured in a colorimetric DNA-binding assay. Double-stranded oligonucleotide containing the p53 consensus DNA-binding sequence was incubated with dilutions of the over-expression lysate and TP53 bound to the oligo was captured onto the surface of a microtiter plate. After washing, bound TP53 was detected with an anti-p53 primary antibody followed by an HRP-labeled secondary antibody. After initial color development, the reaction was quenched and the color intensity was measured at 450nm. Overexpression cell lysates are prepared from HEK293T cells transfected with RC200003 using transfection reagent MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Species: Human
• Expression Host: HEK293T
• Expression cDNA Clone or AA Sequence: TrueORF Clone RC200003
• Tag: C-Myc/DDK
• Detection Antibodies: Clone OTI4C5, Anti-DDK (FLAG) monoclonal antibody (TA50011-100)
• Accession Number: NM_000546, NP_000537
• Other Names: BCC7; BMFS5; LFS1; P53; TRP53
• Predicted MW: 43.7 kDa
• Components: 1 vial of 20 ug lyophilized gene specific transient over-expression cell lysate
• Storage: The lysate can be shipped at ambient temperature. Upon receiving, store the sample at -20°C. Lysate samples can be reconstituted with SDS Sample Buffer. Avoid repeated freeze-thaw cycles after reconstitution. Lysate samples are stable for 12 months from date of receipt when stored at -20°C.
• Preparation: HEK293T cells in 10-cm dishes were transiently transfected with MegaTran Transfection Reagent (TT200002) and 5ug TrueORF cDNA plasmid. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix (Sigma), 1mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate. Protein concentration was measured by BCA kit (Thermo Scientific Inc.). To facilitate transportation and protein, the products are supplied as lyophilized proteins.

Reference Data

• RefSeq: NP_000537
• Locus ID: 7157
• Cytogenetics: 17p13.1
• Protein Families: Druggable Genome, Stem cell - Pluripotency, Transcription Factors
• Protein Pathways: Amyotrophic lateral sclerosis (ALS), Apoptosis, Basal cell carcinoma, Bladder cancer, Cell cycle, Chronic myeloid leukemia, Colorectal cancer, Endometrial cancer, Glioma, Huntington's disease, MAPK signaling pathway, Melanoma, Neurotrophin signaling pathway, Non-small cell lung cancer, p53 signaling pathway, Pancreatic cancer, Pathways in cancer, Prostate cancer, Small cell lung cancer, Thyroid cancer, Wnt signaling pathway