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Human P63 ELISA Kit

SKU:
HUFI02916
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9H3D4
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
TP63, EEC3, KET, OFC8, p40, p51, p73L, SHFM4, TP73L, amplified in squamous cell carcinoma, Chronic ulceRative stomatitis protein, CUSP, EEC3B, p51B, KeRatinocyte transcription factor KET, KETRHS, LMS, OFC8SHFM4TP73LNBP, p40, p51tumor protein p53-like
Reactivity:
Human
Research Area:
Cell Death
€599
Frequently bought together:

Description

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Human P63 ELISA Kit

The Human p63 ELISA Kit is a reliable tool for analyzing levels of the p63 protein in human serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit provides accurate and reproducible results, making it perfect for a variety of research applications.p63 is a key transcription factor that plays a critical role in regulating cell proliferation, differentiation, and apoptosis. Dysregulation of p63 has been linked to various cancers and developmental disorders, making it a valuable biomarker for studying these conditions and developing targeted therapies.

By using the Human p63 ELISA Kit, researchers can gain valuable insights into the role of p63 in disease progression and identify potential therapeutic targets. Trust in this kit for dependable and precise measurement of p63 levels in human samples.

Product Name:Human P63 ELISA Kit
Product Code:HUFI02916
Size:96 Assays
Alias:TP63, EEC3, KET, OFC8, p40, p51, p73L, SHFM4, TP73L, amplified in squamous cell carcinoma, Chronic ulceRative stomatitis protein, CUSP, EEC3B, p51B, KeRatinocyte transcription factor KET, KETRHS, LMS, OFC8SHFM4TP73LNBP, p40, p51tumor protein p53-like, p53CP, P63, p63AIS, p73H, P73H, P73L, p73LB, p51A, TP53CP, TP53L, Transformation-related protein 63, tumor protein p53-competing protein, tumor protein p63, tumor protein p63 deltaN isoform delta, Tumor protein p73-like
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human TP63 concentrations in serum plasma and other biological fluids.
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human TP63 and the recovery rates were calculated by comparing the measured value to the expected amount of Human TP63 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10597
EDTA plasma(n=5)85-10093
UFH plasma(n=5)86-10192
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human TP63 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-103%99-105%86-100%
EDTA plasma(n=5)89-97%83-95%83-93%
UFH plasma(n=5)86-100%84-96%85-97%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ9H3D4
UniProt Protein Function:Function: Acts as a sequence specific DNA binding transcriptional activator or repressor. The isoforms contain a varying set of transactivation and auto-regulating transactivation inhibiting domains thus showing an isoform specificactivity. Isoform 2 activates RIPK4 transcription. May be required in conjunction with TP73/p73 for initiation of p53/TP53 dependent apoptosis in response to genotoxic insults and the presence of activated oncogenes. Involved in Notch signaling by probably inducing JAG1 and JAG2. Plays a role in the regulation of epithelial morphogenesis. The ratio of DeltaN-type and TA*-type isoforms may govern the maintenance of epithelial stem cell compartments and regulate the initiation of epithelial stratification from the undifferentiated embryonal ectoderm. Required for limb formation from the apical ectodermal ridge. Activates transcription of the p21 promoter. Ref.3 Ref.15 Ref.17 Ref.18 Ref.19 Ref.23 Ref.24
UniProt Protein Details:Cofactor: Binds 1 zinc ion per subunit

By similarity.

Subunit structure: Binds DNA as a homotetramer. Isoform compositionof the tetramer may determine transactivation activity. Isoforms Alpha and Gamma interact with HIPK2. Interacts with SSRP1, leading to stimulate coactivator activity. Isoform 1 and isoform 2 interact with WWP1. Interacts with PDS5A. Isoform 5 (via activation domain) interacts with NOC2L. Ref.10 Ref.13 Ref.16 Ref.19 Ref.21 Ref.22 Ref.23

Subcellular location: Nucleus Ref.17 Ref.23.

Tissue specificity: Widely expressed, notably in heart, kidney, placenta, prostate, skeletal muscle, testis and thymus, although the precise isoform variesaccording to tissue type. Progenitor cell layers of skin, breast, eye and prostate express high levels of DeltaN-type isoforms. Isoform 10 is predominantly expressed in skin squamous cell carcinomas, but not in normal skin tissues. Ref.3 Ref.8 Ref.14

Domain: The transactivation inhibitory domain (TID) can interact with, and inhibit the activity of the N-terminal transcriptional activation domain of TA*-type isoforms. Ref.17 Ref.18

Post-translational modification: May be sumoylated

By similarity.Ubiquitinated. Polyubiquitination involves WWP1 and leads to proteasomal degradation of this protein. Ref.21

Involvement inDisease: Acro-dermato-ungual-lacrimal-tooth syndrome (ADULT syndrome) [MIM:103285]: A form of ectodermal dysplasia. Ectodermal dysplasia defines a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. ADULT syndrome involves ectrodactyly, syndactyly, finger- and toenail dysplasia, hypoplastic breasts and nipples, intensive freckling, lacrimal duct atresia, frontal alopecia, primary hypodontia and loss of permanent teeth. ADULT syndrome differs significantly from EEC3 syndrome by the absence of facial clefting.Note: The disease is caused by mutations affecting the gene represented in this entry.Ankyloblepharon-ectodermal defects-cleft lip/palate (AEC) [MIM:106260]: An autosomal dominant condition characterized by congenital ectodermal dysplasia with coarse, wiry, sparse hair, dystrophic nails, slight hypohidrosis, scalp infections, ankyloblepharon filiform adnatum, maxillary hypoplasia, hypodontia and cleft lip/palate.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.29Ectrodactyly, ectodermal dysplasia, and cleft lip/palate syndrome 3 (EEC3) [MIM:604292]: A form of ectodermal dysplasia, a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. It is an autosomal dominant syndrome characterized by ectrodactyly of hands and feet, ectodermal dysplasia and facial clefting.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.26 Ref.27 Ref.30 Ref.32Split-hand/foot malformation 4 (SHFM4) [MIM:605289]: A limb malformation involving the central rays of the autopod and presenting with syndactyly, median clefts of the hands and feet, and aplasia and/or hypoplasia of the phalanges, metacarpals, and metatarsals. Some patients have been found to have mental retardation, ectodermal and craniofacial findings, and orofacial clefting.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.27 Ref.30Limb-mammary syndrome (LMS) [MIM:603543]: Characterized by ectrodactyly, cleft palate and mammary-gland abnormalities.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.30Defects in TP63 are a cause of cervical, colon, head and neck, lung and ovarian cancers.Ectodermal dysplasia, Rapp-Hodgkin type (EDRH) [MIM:129400]: A form of ectodermal dysplasia, a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. Characterized by the combination of anhidrotic ectodermal dysplasia, cleft lip, and cleft palate. The clinical syndrome is comprised of a characteristic facies (narrow nose and small mouth), wiry, slow-growing, and uncombable hair, sparse eyelashes and eyebrows, obstructed lacrimal puncta/epiphora, bilateral stenosis of external auditory canals, microsomia, hypodontia, cone-shaped incisors, enamel hypoplasia, dystrophic nails, and cleft lip/cleft palate.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.33 Ref.34 Ref.35 Ref.36Non-syndromic orofacial cleft 8 (OFC8) [MIM:129400]: A birth defect consisting of cleft lips with or without cleft palate. Cleft lips are associated with cleft palate in two-third of cases. A cleft lip can occur on one or both sides and range in severity from a simple notch in the upper lip to a complete opening in the lip extending into the floor of the nostril and involving the upper gum.Note: The disease is caused by mutations affecting the gene represented in this entry.

Sequence similarities: Belongs to the p53 family.Contains 1 SAM (sterile alpha motif) domain.

Sequence caution: The sequence AAF43486.1 differs from that shown. Reason: Erroneous initiation. The sequence AAF43487.1 differs from that shown. Reason: Erroneous initiation. The sequence AAF43488.1 differs from that shown. Reason: Erroneous initiation. The sequence AAF43489.1 differs from that shown. Reason: Erroneous initiation. The sequence AAF61624.1 differs from that shown. Reason: Frameshift at position 26. The sequence BAA32592.1 differs from that shown. Reason: Frameshift at position 26. The sequence BAA32593.1 differs from that shown. Reason: Frameshift at position 26.

NCBI Summary:This gene encodes a member of the p53 family of transcription factors. An animal model, p63 -/- mice, has been useful in defining the role this protein plays in the development and maintenance of stratified epithelial tissues. p63 -/- mice have several developmental defects which include the lack of limbs and other tissues, such as teeth and mammary glands, which develop as a result of interactions between mesenchyme and epithelium. Mutations in this gene are associated with ectodermal dysplasia, and cleft lip/palate syndrome 3 (EEC3); split-hand/foot malformation 4 (SHFM4); ankyloblepharon-ectodermal defects-cleft lip/palate; ADULT syndrome (acro-dermato-ungual-lacrimal-tooth); limb-mammary syndrome; Rap-Hodgkin syndrome (RHS); and orofacial cleft 8. Both alternative splicing and the use of alternative promoters results in multiple transcript variants encoding different proteins. Many transcripts encoding different proteins have been reported but the biological validity and the full-length nature of these variants have not been determined. [provided by RefSeq, Jul 2008]
UniProt Code:Q9H3D4
NCBI GenInfo Identifier:57013009
NCBI Gene ID:8626
NCBI Accession:Q9H3D4.1
UniProt Secondary Accession:Q9H3D4,O75080, O75195, O75922, O76078, Q6VEG2, Q6VEG3 Q6VEG4, Q6VFJ1, Q6VFJ2, Q6VFJ3, Q6VH20,
UniProt Related Accession:Q9H3D4
Molecular Weight:
NCBI Full Name:Tumor protein 63
NCBI Synonym Full Names:tumor protein p63
NCBI Official Symbol:TP63
NCBI Official Synonym Symbols:AIS; KET; LMS; NBP; RHS; p40; p51; p63; EEC3; OFC8; p73H; p73L; SHFM4; TP53L; TP73L; p53CP; TP53CP; B(p51A); B(p51B)
NCBI Protein Information:tumor protein 63; CUSP; transformation-related protein 63; tumor protein p53-competing protein; amplified in squamous cell carcinoma; chronic ulcerative stomatitis protein; keratinocyte transcription factor KET; tumor protein p63 deltaN isoform delta
UniProt Protein Name:Tumor protein 63
UniProt Synonym Protein Names:Chronic ulcerative stomatitis protein; CUSP; Keratinocyte transcription factor KET; Transformation-related protein 63; TP63; Tumor protein p73-like; p73L; p40; p51
Protein Family:
UniProt Gene Name:TP63
UniProt Entry Name:P63_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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