Anti-Ribosomal P IgG ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

Anti-Ribosomal P (lgG) ELISAis a solid phase enzyme immunoassay employing native human ribosomal P-proteins P0, P1 and P2 isolated from eukaryotic cell line for the quantitative and qualitative detection of antibodies against ribosomal P-proteins(rib-P) in human serum.
The specificity of anti-rib-P antibodies is restricted to a common antigenic determinant located on the highly conserved carboxyl-terminal portion of the three P proteins. The assay is a tool for researching diagnosis of systemic lupus erythematosus (SLE).

Contents of Kit

1. Sample Buffer (5×): 1 × 20 mL. 5× concentrated Tris, sodium chloride (NaCl), bovine serum albumin (BSA), sodium azide < 0.1% (preservative)
2. Wash Buffer (50×): 1 × 20 mL. 50× concentrated Tris, NaCl, Tween 20, sodium azide < 0.1% (preservative)
3. Negative Control: 1 × 1.5 mL. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative)
4. Positive Control: 1 × 1.5 mL. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative)
5. Cut-Off Calibrator: 1 × 1.5 mL. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative)
6. Calibrators: 6 × 1.5 mL. Concentration of each calibrator: 0, 3, 10, 30, 100, 300 U/mL. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative)
7. Conjugate, lgG: 1 × 15 mL. Containing: Antihuman immunoglobuilns conjugated to horseradish peroxidase, bovine serum albumin (BSA)
8. TMB Substrate: 1 × 15 mL. Stabilized tetramethylbenzidine and hydrogen peroxide (TMB/H₂O₂)
9. Stop Solution: 1 × 15 mL. 1M Hydrochloric Acid
10. Microtiter plate: 12 × 8 well strips. With breakaway microwells. Refer to paragraph 1 for coating.

Storage

Store all reagents and the microplate at 2-8°C/35-46°F, in their original containers. Once prepared, reconstituted solutions are stable at 2-8°C/35-46°F for 1 month. Reagents and the microplate shall be used within the expiry date indicated on each component, only. Avoid intense exposure of TMB solution to light. Store microplates in designated foil, including the desiccant, and seal tightly.

Precision

To determine the precision of the assay, the variability (intra- and inter-assay) was assessed by examining its reproducibility on three serum samples selected to represent a range over the tandard curve.

Sensitivity

Testing sample buffer 30 times, Anti-Ribosomal P (IgG) ELISA gave an analytical sensitivity of 1.0 U/mL.

General Description

The ribosomal phosphoproteins Po (~38 kDa), P1(~ 19 kDa) and P2(~17 kDa) are located within the 6os subunit of human ribosomes. In contrast to the majority of basic ribosomal proteins, P1 and P2 are acidic. The ribosomal proteins are assoclated to a pentamer with two P1/P2 heterodimers anchored to Po by the amino terminal porton of P2.This pentamer is located in a highly accessible region on the stalk of the ribosome. Biochemical studies suggest that P1/P2 play a fundamental role in all three phases of ribosomal polypeptide synthesis (initiation translocation, termination).
Research indicates that autoantibodies to ribosomal proteins are highly specific for SLE since they are not found in other autoinmune diseases or in infections. The frequency of anti-rib-P antibodies is 10-20% in randomly selected SLE subjects. Anti-rib-P antibodies are detected more frequently in lupus subjects with severe psychiatric manifestations. In addition, studies suggest that other organ involvement including renal and hepatic disease might be correlated with the presence of anti-rib-P.
Technical Data
Sample Material: serum
Sample Volume: 10 μL of sample diluted 1:101 with 1x sample buffer
Total Incubation Time: 90 minutes at 20-32°C/68-8936°F
Calibration Range: 0-300 U/mL
Analytical Sensitivity: 1.0 U/mL
Storage: at 2-8°C/35-46°F, use original vials only
Number of Determinations: 96 Tests