| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | VB12 |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Mouse Vitamin B12 (VB12) ELISA Kit has high sensitivity and excellent specificity for detection of Mouse VB12. No significant cross-reactivity or interference between Mouse VB12 and analogues was observed.
Background
Cyanocobalamin is the most common and widely produced of the chemical compounds that have vitamin activity as vitamin B12. Vitamin B12 is the "generic descriptor" name for any of such vitamers of vitamin B12. Because the body can[citation needed] convert cyanocobalamin to any one of the active vitamin B12 compounds, by definition this makes cyanocobalamin itself a form (or vitamer) of B12, albeit a largely artificial one. Cyanocobalamin usually does not occur in living organisms, but animals can convert commercially produced cyanocobalamin into active (cofactor) forms of the vitamin, such as methylcobalamin. The amount of cyanide liberated in this process is so small that its toxicity is negligible. Cyanocobalamin is the most famous and widely produced vitamer in the vitamin B12 family (the family of chemicals that function as B12 when put into the body), because cyanocobalamin is the most air-stable of the B12 forms.
This Vitamin B12 (VB12) ELISA kit is validated for use with Cell culture supernatants, Serum, Plasma, Other biological fluids. Samples should be collected, processed, and stored correctly to preserve analyte integrity — avoid repeated freeze-thaw cycles and centrifuge to remove particulates before use. Dilute samples exceeding the kit's detection range using the supplied assay diluent. Hemolytic, icteric, or lipemic samples may affect assay performance and should be tested with caution.
This is a sandwich ELISA kit employing an HRP (horseradish peroxidase)-conjugated secondary antibody paired with a TMB (3,3′,5,5′-tetramethylbenzidine) colorimetric substrate. In the sandwich format, the target analyte Vitamin B12 (VB12) captured on the microplate surface is detected by the conjugated antibody, generating a colorimetric signal proportional to analyte concentration. The reaction is stopped and absorbance measured at 450 nm on a standard microplate reader.
The complete protocol, from sample addition to final plate reading, requires approximately 3–5 hours. This includes two incubation periods (analyte binding and detection antibody steps), intermediate wash cycles to remove unbound material, 15–30 minutes of TMB substrate development, and final stop-solution addition before absorbance reading. Exact timing will vary with experience level and the number of samples processed in parallel.
Required equipment: (1) a microplate spectrophotometer capable of reading absorbance at 450 nm (reference wavelength 540–570 nm recommended for background correction); (2) precision single-channel or multichannel pipettes; (3) a plate washer or multichannel aspirator; (4) a microcentrifuge for sample clarification; and (5) a 37°C incubator or stable room-temperature environment. No fluorescence or luminescence reader is required — standard colorimetric plate readers are fully compatible with this kit.
This ELISA kit is formally validated for Mouse. Cross-reactivity with species not listed in the specification has not been independently characterized. Variability in protein sequence homology across species means that performance in unlisted species cannot be guaranteed without additional validation. For cross-species detection requirements or non-standard sample matrices, please contact BioHippo support or refer to the manufacturer's technical team for guidance.
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Author:Muñoz-del-Carpio-Toia, Agueda, et al. Publication name:HIV Nursing IF:2.3