|Sample type||Average %||Range|
|Cell culture media||90.1||88.3% - 93.4%|
Abcam’s C-Reactive Protein (CRP) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CRP protein in Human cell culture supernatant, serum and plasma 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.
CRP displays several functions associated with host defense: it promotes agglutination, bacterial capsular swelling, phagocytosis and complement fixation through its calcium-dependent binding to phosphorylcholine. CRP can interact with DNA and histones and it may scavenge nuclear material released from damaged circulating cells. CRP is secreted; it forms a homopentamer pentaxin (or pentraxin) which have a discoid arrangement of 5 non-covalently bound subunits. CRP binds 2 calcium ions per subunit. The concentration of CRP in plasma increases greatly during acute phase response to tissue injury, infection or other inflammatory stimuli. It is induced by IL1/interleukin-1 and IL6//interleukin-6.
|Components||1 x 96 tests|
|10X CRP Capture Antibody||1 x 600µl|
|10X CRP Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|CRP Human Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS (ab193972)||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab181416 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.|
Background-subtracted data values (mean +/- SD, n=2) are graphed.
Background subtracted data values (mean +/- SD, n=2) are graphed.
10,000X diluted sera from 10 apparently healthy male donors were measured using this kit. Interpolated data values corrected for sample dilution are graphed in ng of CRP per mL of serum (mean +/- SD, n=2). Nine out of ten individual Human sera tested within or at reported Human serum range (< 5,000 ng/mL, dotted line). The mean of CRP concentration of these nine individual sera was determined to be 2,240 ng/mL with a range of 559 – 5,285 ng/mL. Note that one individual Human serum sample (donor # 2) tested substantially higher, 11,737 ng/mL.
Background subtracted data values of two serum dilutions (as indicated in parenthesis) are graphed (mean +/- SD, n=2).
The concentrations of CRP were interpolated from data values shown above using the CRP standard curve, corrected for sample dilution, and graphed in ng of CRP per mL of serum. As expected, note that the CRP serum concentrations are increased in RA patients.