HIV1 p24 ELISA Kit (ab218268)
- Datasheet
- References (3)
- Protocols
Overview
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Product name
HIV1 p24 ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 3 4.5% Inter-assay Sample n Mean SD CV% Overall 5 4.7% -
Sample type
Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
1.1 pg/ml -
Range
4.69 pg/ml - 300 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 105 94% - 117% Cell culture extracts 107 103% - 113% Cell culture media 102 94% - 107% Hep Plasma 102 100% - 103% EDTA Plasma 98 93% - 103% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
HIV1 p24 in vitro SimpleStep ELISA® kit is designed for the quantitative measurement of HIV1 p24 protein in human serum, plasma, cell culture supernatant, and cell and tissue extract samples.
SimpleStep ELISA® technology 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, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.
ASSAY SPECIFICITY This kit recognizes both native and recombinant HIV-1 p24 protein in serum, EDTA and heparin plasma, cell culture supernatant, and cell and tissue extract samples only. Urine, saliva, and milk samples have not been tested with this kit.
CROSS REACTIVITY Recombinant HIV-1 Gag protein was prepared at 50 ng/mL and 1 ng/mL and assayed for cross reactivity. 1% cross-reactivity was observed.
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Notes
HIV1 p24 (capsid) protein is essential for HIV-1 viral replication and for the HIV-1 infection of non-dividing cells. HIV1 p24 proteins form viral capsid that encapsulates the genomic HIV1 RNA. HIV1 p24 concentration in host plasma is commonly used as indicator of viral load. Upon the viral infection, the development of anti-HIV1 p24 host humoral responses leads to immune complex formation and reduction of the free HIV1 p24 concentration in circulation.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X HIV1 p24 Capture Antibody 1 x 600µl 10X HIV1 p24 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml HIV1 p24 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent 50BS 1 x 20ml Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Relevance
HIV1 performs highly complex orchestrated tasks during the assembly, budding, maturation and infection stages of the viral replication cycle. During viral assembly, the proteins form membrane associations and self-associations that ultimately result in budding of an immature virion from the infected cell. Gag precursors also function during viral assembly to selectively bind and package two plus strands of genomic RNA. Capsid protein p24 probably forms the conical core of the virus that encapsulates the genomic RNA-nucleocapsid complex. -
Cellular localization
Membrane -
Alternative names
- CA
- Capsid protein p24
- HIV-1 Gag p24
see all
Images
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Other - HIV1 p24 ELISA Kit (ab218268)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 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.
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Example of HIV1 p24 standard curve in Sample Diluent 50BSBackground-subtracted data values (mean +/- SD) are graphed.
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Raw data values for Example of HIV-1 p24 standard curve in Sample Diluent 50BS.Raw data values are shown in the table
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Example of HIV1 p24 standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Raw data values for example of HIV-1 p24 standard curve in Sample Diluent NS.Raw data values are shown in the table
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Example of HIV1 p24 standard curve in 1X Cell Extraction Buffer PTRBackground-subtracted data values (mean +/- SD) are graphed.
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Example of HIV-1 p24 standard curve in 1X Cell Extraction Buffer PTRRaw data values are shown in the table
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Interpolated concentrations of spike HIV1 p24 in human serum, human plasmas, and rhesus macaque plasmaThe concentrations of HIV1 p24 were measured in duplicates, interpolated from the HIV1 p24 standard curves and corrected for sample dilution. Undiluted samples are as follows: human serum 100% (neat), human plasma (heparin) 100% (neat), human plasma (EDTA) 100% (neat), and rhesus macaque plasma (EDTA) 100% (neat). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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Interpolated concentrations of spike HIV1 p24 in Jurkat Cell Extract and RPMI + 10% FBS cell culture mediaThe concentrations of HIV1 p24 were measured in duplicates, interpolated from the HIV1 p24 standard curves and corrected for sample dilution. Undiluted samples are as follows: Jurkat cell extract 100 µg/mL, RPMI + 10% FBS cell culture media 100% (neat). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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Serial dilutions of recombinant HIV1 p24 were prepared within the working range of the assay and assayed for reactivity.Serial dilutions of recombinant HIV1 p24 (group M, subtype B, strain 92418), HIV1 p24 (group M, subtype C, strain 92BR025) and HIV1 p24 (group O, strain BCF06) were prepared within the working range of the assay and assayed for reactivity. O.D. values within the linear range of each protein are graphed.
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Acid treatment recoveryThree concentrations of purified HIV-1 p24 protein were spiked in duplicate to the indicated biological matrix and treated with the Acid Treatment Protocol to evaluate signal recovery in the working range of the assay. The signals of the same concentrations of purified HIV-1 p24 protein spiked in duplicate to Sample Diluent 50BS and treated with the Acid Treatment Protocol were taken as 100%.
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Acid treatment effectSingle concentrations of purified HIV-1 p24 protein were spiked in duplicate to the indicated biological matrix and treated with the Acid Treatment Protocol or mock acid treated to evaluate the effect of the Acid Treatment Protocol on the signal. The signals of single concentrations of purified HIV-1 p24 protein spiked in duplicate to the indicated biological matrix and mock acid treated were taken as 100%.
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Acid treatment recovery of synthetic complexesSingle concentrations of purified HIV-1 p24 protein were spiked to the indicated biological matrix, pre-incubated with unlabeled capture and detector antibodies or unrelated antibodies and treated with the Acid Treatment Protocol to evaluate the signal recovery of p24 complexed with interfering antibodies by Acid Treatment Protocol. The signals of single concentrations of purified HIV-1 p24 protein spiked to the indicated biological matrix, preincubated with unrelated antibodies and treated with the Acid Treatment Protocol were taken as 100%. There were no signals if the single concentrations of purified HIV-1 p24 protein were spiked to the indicated biological matrix, pre-incubated with the unlabeled capture and detector antibodies and mock acid treated, indicating that the unlabeled capture and detector antibodies efficiently formed synthetic immune complexes with HIV-1 p24 and completely blocked the signals.
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Linearity of dilution – spiked Purified HIV-1 p24 in Human serum, plasma (heparin, citrate, EDTA) and rhesus macaque EDTA plasma.Purified HIV-1 p24 was spiked into biological samples and diluted in a 2-fold dilution series in Sample Diluent 50BS.
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Linearity of dilution – spiked purified HIV-1 p24 in FBS cell culture media and Jurkat Cell ExtractPurified HIV-1 p24 was spiked into biological samples and diluted in a 2-fold dilution series in Sample Diluent NS for cell culture media and 1X Cell Extraction Buffer PTR for cell extract samples.
Protocols
References
This product has been referenced in:
- Ma X et al. TRIM28 promotes HIV-1 latency by SUMOylating CDK9 and inhibiting P-TEFb. Elife 8:N/A (2019). Read more (PubMed: 30652970) »
- Pan T et al. USP49 potently stabilizes APOBEC3G protein by removing ubiquitin and inhibits HIV-1 replication. Elife 8:N/A (2019). ELISA ; Human . Read more (PubMed: 31397674) »
