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AB325617

Human p53 ELISA Kit, Chemiluminescent

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Human p53 ELISA Kit, Chemiluminescent is a 55 min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human p53 with higher sensitivity and broader measuring range.

- Single 30 min incubation and single -wash step for results in less than an hour
- Chemiluminescent detection allows a broad dynamic range : 0.02 - 50 ng/ml with a sensitivity of 0.009 ng/mL
- Patent pending

View Alternative Names

P53, TP53, Cellular tumor antigen p53, Antigen NY-CO-13, Phosphoprotein p53, Tumor suppressor p53

6 Images
Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

Standard curve comparison between the Chemiluminescent Human p53 SimpleStep ELISA and colorimetric Human p53 SimpleStep ELISA (ab171571) in 1X Cell Extraction Buffer PTR. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

Dynamic range comparison between the Chemiluminescent Human p53 SimpleStep ELISA, colorimetric Human p53 SimpleStep ELISA (ab171571), and leading colorimetric competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity and increased dynamic range.

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

Interpolated concentration of native p53 was measured in duplicate at different sample concentrations. Undiluted samples are as follows : SH-SY5Y extract 31.25 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

Comparison of p53 expression in different cell lines. Background subtracted data from duplicate measurements are plotted. Note the relative lower expression of p53 in the MCF-7, HeLa and HL-60 cell lines compared to HEK293 and SH-SY5Y.

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)
  • sELISA

Lab

Sandwich ELISA - Human p53 ELISA Kit, Chemiluminescent (AB325617)

Comparison of p53 expression in different cell lines by ELISA (barchart) and western blot (top). Raw OD450 nm data from duplicate measurements for the indicated cells lines is shown (20 ug lysate analyzed). The p53 capture antibody (ab154036) was used to blot the same lysates as analyzed by ELISA (20 ug loaded/lane). The GAPDH blot is included to show the relative loads of each lysate.

Key facts

Detection method

Luminescent

Sample types

Cell Extract, Tissue Extracts

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 0.009 ng/mL

Range

0.02 - 50 ng/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human p53 ELISA Kit, Chemiluminescent (ab325617) is a rapid single-wash sandwich 55 min ELISA to measure Human p53 in Cell Extract, Tissue Extract with a sensitivity of 0.009 ng/mL.

How the assay works

Human p53 ELISA Kit, Chemiluminescent (ab325617) SimpleStep Ignite™ ELISA employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colormetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human p53 (ab325617) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 4.3" }, { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 6.7" } ]
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Recovery

[ { "sample": "Cell Lysate", "range": "93 - 107 %", "average": "= 99" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab325617-1x96Tests", "assetComponentDetails": [ { "size":"1 x 6 mL", "name":"4X Antibody Diluent EB", "number":"AB325617-CMP04", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human p53 Lyophilized Recombinant Protein", "number":"AB325617-CMP03", "productcode":"" }, { "size":"1 x 3 mL", "name":"ChemiHRP Reagent B", "number":"AB325617-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human p53 Detector Antibody", "number":"AB325617-CMP02", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seal", "number":"AB325617-CMP11", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated Black 96-Well Microplate", "number":"AB325617-CMP10", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB325617-CMP06", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human p53 Capture Antibody", "number":"AB325617-CMP01", "productcode":"" }, { "size":"1 x 3 mL", "name":"ChemiHRP Reagent A", "number":"AB325617-CMP08", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB325617-CMP07", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB325617-CMP05", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

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Target data

Multifunctional transcription factor that induces cell cycle arrest, DNA repair or apoptosis upon binding to its target DNA sequence (PubMed : 11025664, PubMed : 12524540, PubMed : 12810724, PubMed : 15186775, PubMed : 15340061, PubMed : 17317671, PubMed : 17349958, PubMed : 19556538, PubMed : 20673990, PubMed : 20959462, PubMed : 22726440, PubMed : 24051492, PubMed : 24652652, PubMed : 35618207, PubMed : 36634798, PubMed : 38653238, PubMed : 9840937). Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type (PubMed : 11025664, PubMed : 12524540, PubMed : 12810724, PubMed : 15186775, PubMed : 15340061, PubMed : 17189187, PubMed : 17317671, PubMed : 17349958, PubMed : 19556538, PubMed : 20673990, PubMed : 20959462, PubMed : 22726440, PubMed : 24051492, PubMed : 24652652, PubMed : 38653238, PubMed : 9840937). Negatively regulates cell division by controlling expression of a set of genes required for this process (PubMed : 11025664, PubMed : 12524540, PubMed : 12810724, PubMed : 15186775, PubMed : 15340061, PubMed : 17317671, PubMed : 17349958, PubMed : 19556538, PubMed : 20673990, PubMed : 20959462, PubMed : 22726440, PubMed : 24051492, PubMed : 24652652, PubMed : 9840937). One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression (PubMed : 12524540, PubMed : 17189187). Its pro-apoptotic activity is activated via its interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 (PubMed : 12524540). However, this activity is inhibited when the interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 is displaced by PPP1R13L/iASPP (PubMed : 12524540). In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seems to have an effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. Regulates the circadian clock by repressing CLOCK-BMAL1-mediated transcriptional activation of PER2 (PubMed : 24051492).
See full target information TP53
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websiteProtocolBooklet
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Product promise

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