Overview
Since 7.4 is the typical pH of human blood, many diagnostic assays are created at this level.
However, it is necessary to differentiate between coating or spotting buffers and assay buffers for solid phase assays, such as microarray applications. These two process steps have different buffer specifications.
Maximum immobilization effectiveness, uniform spot morphology, and upstream assay requirements are all met while the capture biomolecule is kept functional.
Moreover, it is not just a matter of pH; if the immobilization chemistry relies on the coupling of primary amines to functional groups on the surface, buffers with amines like Tris must be avoided. The involved biomolecules must function as well as possible in the assay buffer.
For instance, with a 10-fold dilution, the pH of a 100 mM sodium phosphate buffer rises from 6.7 to 6.9.
In most circumstances, a concentration of 25–50 mM is required to ensure a sufficient buffering capacity. The user could try a lower concentration, such as 10 mM, if the assay uses enzymes that are sensitive to high ionic strength.
Image Credit: Scienion
Applications
- sciBUFFERs for DNA and Oligos
Image Credit: Scienion
Table 1. Source: Scienion
| Product Name |
Product Description |
Contains |
Product Number |
| sciSPOT cDNA |
Printing buffer based on SSC (pH 7) with a detergent, to enable optimal microarray printing of cDNA samples. |
50 mL 2x conc. |
CBD-5411-50 |
| sciSPOT Oligo B1 |
Printing buffer based on SSC (pH 7) and is optimized for use with oligonucleotides on functionalized glass slides. |
50 mL 2x conc. |
CBD-5421-50 |
| sciSPOT Oligo B2 |
Printing buffer based on phosphate buffer (pH 9), for producing microarrays on polymer surfaces (sciPLEXPLATEs) |
50 mL 2x conc. |
CBD-5418-50 |
| sciPROCESS-cDNA |
Blocking buffers for post-printing treatment, used to deactivate the residual functional groups on the surface and minimize unspecific binding of the target DNA/RNA. |
500 mL 2x conc. |
CBD-5416-500 |
| sciPROCESS-Oligo |
|
500 mL 2x conc. |
CBD-5426-500 |
| sciPROCESS-Oligo |
|
300 mL 2 x conc. |
CBD-5426-300 |
| sciHYB |
Hybridization buffer to labeled cDNA or PCR products and immobilized capture probes, while minimizing cross-hybridization. Hybridization is accelerated and more efficient due to minimized secondary structures in probes and target sequences even at lower temperatures (42 °C). |
1.5 mL 1x conc. |
CBD-5417-1.5 |
| sciHYB M1 |
Hybridization buffer, that, in contrast to standard sciHYB, consists of a non-toxic formulation and is used at more elevated temperatures (55-65 °C). |
50 mL 1 x conc. |
CBD-5419-50 |
| sciWASH I DNA |
Washing buffers to reduce background signal after the hybridization step. Contain a detergent with ionic strengths for different levels of stringency. |
500 mL 5x conc. |
CBD-5413-500 |
| sciWASH II DNA |
|
500 mL 5x conc. |
CBD-5414-500 |
| sciWASH III DNA |
|
500 mL 5x conc. |
CBD-5415-500 |
| sciCOLOR T2 |
HRP substrate purpose-built for microarray applications |
50 mL |
CD-5600-50 |
| 100 mL |
CD-5600-100 |
| 250 mL |
CD-5600-250 |
| sciCOLOR T3 |
Substrate purpose-built for black staining microarray applications |
50 mL |
CD-5603-50 |
| 100 mL |
CD-5603-100 |
| 250 mL |
CD-5603-250 |
Image Credit: Scienion
Table 2. Source: Scienion
| Product Name |
Product Description |
Contains |
Product Number |
| sciSPOT Protein D1 |
Printing buffer based on PBS at pH 7.2. PBS is a nontoxic buffer that mimics the osmolarity and ion concentrations of the human body. |
25 mL 2x conc. |
CBP-5431-25 |
| sciSPOT Protein D11 |
Printing buffer based on sciSPOT Protein D1, but contains an additive to slow down evaporation, which improves spot homogeneity. |
25 mL 2x conc. |
CBP-5435-25 |
| sciSPOT Protein D12 |
Printing buffer based on sciSPOT Protein D1, but includes an additive to stabilize proteins, which helps maintaining nativity of the proteins and promotes shelf life of the microarrays. |
25 mL 2x conc. |
CBP-5436-25 |
| sciSPOT Protein D2 |
Sodium Phosphate based buffer at pH 7.5. |
25 mL 2x conc. |
CBP-5439-25 |
| sciSPOT Protein D3 |
Printing buffer based on TBS, pH 8.0. |
25 mL 2x conc. |
CBP-5441-25 |
| sciSPOT Protein D4 |
Carbonate based printing buffer, pH 9.6 |
1.5 mL 1x conc. |
CBP-5442-25 |
| sciSTAB S3 |
Protein stabilizer that can be added to any of our sciSPOT buffers. This will enhance shelf life and functionality of your protein microarrays by stabilizing the native structure of the arrayed proteins. |
2 mL 5 x conc. |
CD-5601-2 |
| sciBLOCK Protein D1 |
Designed for post-print processing of the protein microarrays and will passivate the surface. This PBS based buffer efficiently blocks unspecific binding, resulting in low background. |
500 mL 5x conc. |
CBP-5433-500 |
| sciBLOCK Protein D1M |
|
100 mL 1x conc. |
CBP-5438-100 |
| sciWASH Protein D1 |
PBS based washing buffer, suitable for all stages during multiplex ELISA. |
500 mL 8x conc. |
CBP-5432-500 |
| sciWASH Protein D1M |
|
250 mL 5x conc. |
CBP-5437-250 |
| sciBIND Protein D1 |
PBS based reagent diluent, which is ideally suited for dilution of your samples and secondary antibodies. This buffer is formulated to enhance your signals while reducing cross-reactivity. |
1.6 mL 2x conc. |
CBP-5434-1.6 |
| 50 mL 2x conc. |
CBP-5434-50 |
| sciCOLOR T2 |
HRP substrate purpose-built for microarray applications |
50 mL |
CD-5600-50 |
| 100 mL |
CD-5600-100 |
| 250 mL |
CD-5600-250 |
| sciCOLOR T3 |
Substrate purpose-built for black staining microarray applications |
50 mL |
CD-5603-50 |
| 100 mL |
CD-5603-100 |
| 250 mL |
CD-5603-250 |
| sciSTAB S3 |
Protein stabilizer that can be added to any of our sciSPOT buffers. This will enhance shelf life and functionality of your protein microarrays by stabilizing the native structure of the arrayed proteins. |
2 mL 5x conc. |
CD-5601-2 |
Table 3. Source: Scienion
| Product Name |
Product Description |
Contains |
Product Number |
| sciBUFFERSET Oligo M1 |
Set of buffers for use with oligonucleotides on polymer surfaces |
sciSPOT Oligo B2, sciPROCESS Oligo, sciHYB M1 |
CBP-5450 |
| sciBUFFERSET Protein D1 |
Set of buffers for optimal microarray printing of protein samples |
sciSPOT Protein D1, sciWASH Protein D1, sciBLOCK Protein D1, sciBIND Protein D1 |
CBP-5410 |
| sciBUFFERSET Protein D1M |
Set of buffers for optimal microarray printing of protein samples, up to 5 x96 well plates |
sciSPOT Protein D1, sciWASH Protein D1M, sciBLOCK Protein D1M, sciBIND Protein D1M |
CBP-5440 |
Table 4. Source: Scienion
| Product Name |
Product Description |
Contains |
Product Number |
| sciMULTIPLEX Box |
sciMULTIPLEX system contains selected consumables to facilitate transfer of multiple traditional singleplex immunoassays to one multiplex immunoassay. The intention of this kit is to expedite setting up a proof-of-principle assay as a solid base for further development and validation. |
2x sciSOURCEPLATE 384 PP, 2 sciPLEXPLATE Type 1, 2x Type 2 and 2x solid black, sciSPOT Protein D1, D2, D3, D4 (5 mL each at 2 conc.), sciSTAB S3, sciBLOCK Protein D1M, 2x sciWASH Protein D1 (75 mL, 8x conc.) and sciCOLOR T2 |
CD-5602 |