ab195215 IgG Human SimpleStep ELISA Kit

Product nameIgG Human SimpleStep ELISA? KitDetection methodColorimetricPrecision Intra-assay Sample n Mean SD CV% Serum 8 6.4% Inter-assay Sample n Mean SD CV% Serum 3 14.7% Sample typeCell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Tissue LysateAssay typeSandwich (quantitative)Sensitivity 0.02 ng/mlRange0.23 ng/ml - 15 ng/mlRecovery Sample specific recovery Sample type Average % Range Saliva 89 80% - 106% Milk 89 83% - 94% Urine 87 82% - 93% Serum 101 88% - 125% Cell culture media 107 96% - 115% Heparin Plasma 100 - EDTA Plasma 90.17 87% - 93% Citrate Plasma 100 98% - 102% Assay time1h 30m Assay durationOne step assaySpecies reactivityReacts with: Human Product overview Abcam’s Human IgG in vitro SimpleStep ELISA? (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IgG protein in Human serum, plasma, milk, urine, saliva, culture media and tissue extracts. 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. Notes Immunoglubulin G (IgG) is a glycoprotein molecule which belongs to the immunoglobulin family of proteins known as antibodies. Immunoglobulins are the key component of humoral immunity. IgG has an approximate molecular weight of about 150kDa and it is composed of four peptide chains: two identical heavy chains (γ) of about 50kDa and two identical light chains (κ) of about 25kDa each. The heavy chains are linked to each other and to the light chain by disulfide bonds. At the N terminus, both the heavy and the light chain contain variable regions (VH and VL) which account for antibody diversity. At the C terminus, both chains contain constant regions (CH and CL) but only CH mediates effector functions. Structurally the IgG molecule may be divided into: (1) the Fragment antigen binding region (Fab) containing the VL, VH, CL and CH2 all of which shape the antigen binding site and (2) the Fragment crystallizable region (Fc) containing CH domains 2 – 4 which stabilize the antibody and bind to the Fc receptor on the surface of macrophages, neutrophils, natural killer cells as well as to complement proteins to mediate therefore physiological effects. IgG is synthesized and secreted by plasma B cells in response to an immunogen after recognition of specific epitopes on the antigen and it is generated following class switching and maturation of an antibody response, thus providing immune protection. There are four subclasses of IgG in humans (IgG 1, 2, 3, 4) with variable affinity to Fc receptors and complement. The levels of IgG are generally considered to be indicative of an individual’s immune status and are found increase in all types of infections, liver disease, severe malnutrition, dysproteinemia and rheumatoid arthritis. It is decrease in conditions such as hypogammaglobulinemia, X-linked agammaglobulinemia, lymphoid aplasia and chronic lymphoblastic leukemia. IgG accounts for 75% of the total human protein and can be found in serum, lymphatic fluid, cerebrospinal fluid, colostrum, milk, urine, saliva, sweat and body tissues. IgG has been shown to bind some bacterial strains from cutaneous microbiota. The Fc portion of human IgG is frequently used as the basis of prolonged pharmacokinetics as it is used as a fusion partner to extend the half-life of fusion proteins. Tested applicationsSandwich ELISAmore details PlatformMicroplate Properties Storage instructionsStore at +4°C. Please refer to protocols. Components 1 x 96 tests 10X Human IgG Capture Antibody 1 x 600μl 10X Human IgG Detector Antibody 1 x 600μl 10X Wash Buffer PT 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CP 1 x 6ml IgG Human Lyophilized Purified Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate 1 unit Stop Solution 1 x 12ml TMB Substrate 1 x 12ml Alternative names FLJ39988 FLJ40036 FLJ40253 see allDatabase links Entrez Gene: 3500 Human Entrez Gene: 3501 Human Entrez Gene: 3503 Human Entrez Gene: 3504 Human Omim: 147100 Human Omim: 147120 Human Omim: 147130 Human Omim: 147110 Human see all Applications Our Abpromise guarantee covers the use of ab195215 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. Application Abreviews Notes Sandwich ELISA Use at an assay dependent concentration.