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|Sample type||Average %||Range|
|Cell culture supernatant||86.4||85.4% - 87.4%|
|Urine||91.6||89.9% - 92.6%|
|Serum||79.5||78.5% - 80.5%|
|Citrate Plasma||99.2||94.6% - 102.3%|
Survivin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (ab183361) is designed for the quantitative measurement of Survivin protein in human cell culture supernatant, serum, plasma, urine, cell and tissue extract samples.
The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm.Optionally,instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Samples diluted in Sample Diluent NS - 28 pg/mL
Samples diluted in Sample Diluent 75BP - 7 pg/mL
Samples diluted in 1X Cell Extraction Buffer - 25 pg/mL
Survivin is a multitasking protein that has dual roles in promoting cell proliferation and preventing apoptosis. Survivin is a component of a chromosome passage protein complex (CPC) which is essential for chromosome alignment and segregation during mitosis and cytokinesis. Survivin acts as an important regulator of the localization of this complex; directs CPC movement to different locations from the inner centromere during prometaphase to midbody during cytokinesis and it participates in the organization of the center spindle by associating with polymerized microtubules. Survivin complex with RAN plays a role in mitotic spindle formation by serving as a physical scaffold to help deliver the RAN effector molecule TPX2 to microtubules. Survivin may counteract a default induction of apoptosis in G2/M phase. The acetylated form of Survivin represses STAT3 transactivation of target gene promoters. Survivin may play a role in neoplasia. Survivin is an inhibitor of CASP3 and CASP7. Isoform 2 and isoform 3 of Survivin do not appear to play vital roles in mitosis. Isoform 3 shows a marked reduction in its anti-apoptotic effects when compared with the displayed wild-type isoform.
|Components||1 x 96 tests|
|10X Survivin Capture Antibody||1 x 600µl|
|10X Survivin 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|
|ab221825 - Antibody Diluent 5BI||1 x 6ml|
|Plate Seals||1 unit|
|Sample Diluent 75BP||1 x 20ml|
|Sample Diluent NS (ab193972)||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 x 96 tests|
|Stop Solution||1 x 12ml|
|Survivin Human Lyophilized Recombinant Protein||2 vials|
|TMB Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab183361 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Standard curve comparison between human survivin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.
Figure 1. Example of Survivin standard curve for cell culture supernatant and urine samples measurements. Background-subtracted data values (mean +/- SD) are graphed.
Figure 2. Example of Survivin standard curve for serum and plasma-citrate samples measurements. Background-subtracted data values (mean +/- SD) are graphed.
Figure 3. Example of Survivin standard curve for cell and tissue extract samples measurements. Background-subtracted data values (mean +/- SD) are graphed.
Figure 4. Titration of Jurkat cell extract within the working range of the assay. Background-subtracted data values (mean +/- SD, n=2) are graphed.
Figure 5. Survivin concentrations in 10 individual Human serum donors. Two fold diluted serum samples from 10 apparently healthy male donors were measured using this kit. Interpolated data values corrected for sample dilution are graphed in pg of Survivin per mL of serum (mean +/- SD, n=3).
Figure 6. Comparison of Survivin concentrations in media and MCF7 cell supernatants. MCF7 cells were grown in 10F HGDMEM medium and Survivin concentrations were measured in undiluted cell culture supernatant (SN) sample and the 10F HGDMEM medium using this kit. Interpolated data values are graphed (mean +/- SD, n=2). Note that no detectable Survivin concentrations in growth medium were observed.
Figure 7. Comparison of Survivin concentrations in control and nocodazole treated HeLa cells. HeLa cells were grown in 10F HGDMEM medium for 17 hours in the presence of 200 ng/mL nocodazole or drug’s vehicle (DMSO). Survivin concentrations were measured in cell extracts diluted to 125 and 62.5 µg/mL using this kit. Interpolated data values expressed in pg Survivin per g of cell extract are graphed (mean +/- SD, n=2).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"