Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [2]
- Immunohistochemistry [1]
- Other assay [3]
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- Product number
- PA5-22302 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DPYD Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: Raji, K562, NCI-H929. Predicted reactivity: Mouse (91%), Rat (92%), Xenopus laevis (91%), Pig (94%), Bovine (96%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.59 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references P53 represses pyrimidine catabolic gene dihydropyrimidine dehydrogenase (DPYD) expression in response to thymidylate synthase (TS) targeting.
Gokare P, Finnberg NK, Abbosh PH, Dai J, Murphy ME, El-Deiry WS
Scientific reports 2017 Aug 29;7(1):9711
Scientific reports 2017 Aug 29;7(1):9711
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of DPYD using 30 µg of A) Raji (B) K562 and C) NCI-H929 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a DPYD polyclonal antibody (Product # PA5-22302) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DPYD Polyclonal Antibody (Product # PA5-22302). Whole cell extract (30 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with DPYD Polyclonal Antibody (Product # PA5-22302) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DPYD Polyclonal Antibody detects DPYD protein at cytoplasm on H358 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded H358 xenograft. DPYD Polyclonal Antibody (Product # PA5-22302) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 2 p53-dependent repression of DPYD expression in intact liver and impact of human p53 polymorphic variants on liver expression of DPYD . ( A and B ) Fold-change in expression of DPYD mRNA and protein in livers of p53 +/+ (wild-type,) and p53 -/- mice. P-values are determined Multiple t test.and one way Anova [NT vs 5-FU in p53 +/+ mice for DPYD mRNA is p = 0.08; DPYD protein p = 0.0021and NT vs 5-FU in p53 -/- mice for DPYD mRNA is p = 0.040; DPYD protein p = 0.09] ( C ) HUPKI Codon R72P MEF-R/R or MEF-P/P were treated with 5-FU (384 muM) up to 24 hr and DPYD protein expression in MEF-P/P or MEF-R/R allele was evaluated by western blot. ( D ) DPYD protein expression in Normal human fibroblast cell line harboring Codon R72P polymorphism, i.e., NHF P/P and NHF R/R is evaluated by western blot after treatment with 5-FU for the indicated times.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 The tumor suppressor p53 represses dihydropyrimidine dehydrogenase (DPYD) expression. (A) mRNA and protein expression of DPYD in HCT-116 p53 +/+ and HCT-116 p53 -/- cell lines at indicated times after 5-FU (384 muM) treatment. (B) Fold-expression of mRNA in A549 and H460 cell lines at 24 hr after 5-FU (384 muM) treatment with and without siRNA knockdown of p53 (P = 0.0011 N = 3). (C) H3K9 Acetylation at DPYD promoter following 5-FU treatment for indicated time points. Values are normalized in the sequence [input > IgG > total H3 > No Treatment (NT)] (N = 3). (D , E , F) Protein expression of DPYD in A549, U87MG, HT-1080 and H460 is shown after western blotting at the indicated time points with and without siRNA knockdown of p53. (G) H3K4me3 and H3K27me3 at DPYD promoter at 24hrs after 5-FU treatment. Values normalized in the sequence [input > IgG > total H3] (N = 3).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 TP53 -specific liver depletion upregulates the catabolism of 5-FU through DPYD. (A) . Liver specific expression of Cre in Albcre;mT/mG;p53 Delta/Delta mice as seen by expression of GFP and normal histology of liver, Bone marrow (BM) and colon in these mice (B) 5-FU treatment schedule for mice (p53 Delta/+ and p53 Delta/Delta ) with liver specific deletion of the TP53 gene, [S-1 = First dose Vehicle (6hrs) + second dose 5-FU(30 min)]; S-2 = [First dose 5-FU(6hrs) + second dose 5-FU(30 min)]. (C) Ratio of the amount of 5-FUH 2 /5-FU in plasma of liver specific p53 Delta/ + and p53 Delta/Delta genotypes following the treatment plan described in ( B ) (Values represent median n = 5-7; p = 0.0313 Wilcoxon-rank-sum test). (D) Tumor growth delay (TGD) of syngeneic p53dmc-Ras-Myc colonocytes injected subcutaneously (s.c.) into liver specific p53 Delta/ + and p53 Delta/Delta and treated with vehicle or 5-FU IV (100 mg/kg/week, for a total of 6 weeks) (N = 3-5, Doubling time calculated by exponential growth equation). (E) Analysis of cell death and proliferation in syngeneic tumors on p53 Delta/ + and p53 Delta/Delta at the study in ( C ) as indicated by number of apoptotic and mitotic nu clei(N = 3). (F) Representative western blot showing expression of DPYD in liver of p53 Delta/ + and p53 Delta/Delta mice with prior treatment of 5-FU for 6 hrs as indicated in ( B ) and ( C ).