Integral to the accuracy and sensitivity of ELISA techniques are ELISA reagents, including blocking buffers and substrates. Blocking buffers are crucial for preventing non-specific protein binding on the plate, effectively reducing background noise and enhancing the specificity of the signal. They are indispensable in ensuring that ELISA reagents interact specifically with the target analyte. Substrates, another key category of ELISA reagents, are selected based on the desired detection method, significantly affecting the assay's sensitivity and dynamic range. With decades of expertise in antibody and assay development, Rockland's reagents are engineered to optimize performance across a broad spectrum of ELISA applications.
ELISA Essentials
ELISA Blocking Buffers
Rockland provides a selection of premium ELISA blocking buffers, each formulated to minimize non-specific binding and enhance assay performance. The choice of buffer is guided by the enzyme label and antibodies used in your assay, ensuring optimal performance.
| Product | Application Note | Precautions |
| 10X BBS Fish Gel Concentrate | Fish gel is less likely to cross-react with antibodies of mammalian origin than conventional blocking reagents such as non-fat dry milk and BSA. | |
| 10X PBS Fish Gel Concentrate | Fish gel is less likely to cross-react with antibodies of mammalian origin than conventional blocking reagents such as non-fat dry milk and BSA. | For AP antibody labels use 10X TBS Fish Gel Concentrate or 10X BBS Fish Gel Concentrate buffers instead. |
| 10X TBS Fish Gel Concentrate | Fish gel is less likely to cross-react with antibodies of mammalian origin than conventional blocking reagents such as non-fat dry milk and BSA. | |
| Bovine Serum Albumin 30% Solution | BSA blocking solutions are preferred with biotin and AP antibody labels. | |
| Bovine Serum Albumin - Fraction V | BSA blocking solutions are preferred with biotin and AP antibody labels. | Since BSA is notoriously difficult to dissolve, you can save time using our Bovine Serum Albumin 30% Solution and dilute further. |
| Normal Goat Serum (NGS) | For use in ELISA where the detection antibody was produced in goat. | |
| Normal Horse Serum (NHS) | For use in ELISA where the detection antibody was produced in horse. | |
| Normal Mouse Serum (NMS) | For use in ELISA where the detection antibody was produced in mouse. | |
| Normal Rabbit Serum (NRS) | For use in ELISA where the detection antibody was produced in rabbit. | |
| Normal Rat Serum (NRS) | For use in ELISA where the detection antibody was produced in rat. | |
| Normal Sheep Serum (NSS) | For use in ELISA where the detection antibody was produced in sheep. | |
| Blotto Immunoanalytical Grade (Non-Fat Dry Milk) | Immunoanalytical grade non-fat dry milk. | Not suitable for use with biotin-avidin detection due to inherent biotin content. |
| Blotto A Pre-Mixed | General purpose blocking agent. | |
| Blotto B Pre-Mixed | General purpose blocking agent when phospho-specific antibodies are used. | |
| Blocking Buffer Sampler Kit | Set of different blocking buffers for all sorts of immunoassays. |
ELISA Substrates
In selecting ELISA substrates, both the detection method and enzyme label are critical. Luminescent substrates, used with horseradish peroxidase (HRP), are superior for low-abundance analyte detection, offering high sensitivity and a broad dynamic range. Colorimetric substrates, available for both HRP and alkaline phosphatase (AP), yield soluble colored reaction products, making them suitable for either kinetic (e.g., TMB, ABTS) or endpoint assays (e.g., TMB, ABTS, NPP). This flexibility supports diverse assay requirements, balancing sensitivity with operational simplicity.
| Product | Detection Type | Compatible Label |
| Chemiluminescent FemtoMax™ Super Sensitive HRP Substrate | Luminescent | Peroxidase (HRP) |
| UniGlow™ - One Component Chemiluminescent Substrate | Luminescent | Peroxidase (HRP) |
| ABTS ELISA Peroxidase Substrate | Colorimetric | Peroxidase (HRP) |
| TMB ELISA Peroxidase Substrate | Colorimetric | Peroxidase (HRP) |
| TMB Prestained Red ELISA Peroxidase Substrate | Colorimetric | Peroxidase (HRP) |
| NPP (1X) ELISA Alkaline Phosphatase Substrate | Colorimetric | Alkaline phosphatase (AP) |
| NPP (50X) ELISA Alkaline Phosphatase Substrate | Colorimetric | Alkaline phosphatase (AP) |
