Confirm reagent and antibody information
pH630 Deep Red anti-human labeling reagent (QH-17-1003) is supplied as 200 μL of liquid at 0.5 mg/mL in 10 mM PBS, pH 7.4, with 50% glycerol. Mix gently; no reconstitution or sonication is required. Record antibody species, subclass, intact or fragment format and formulation. The name “anti-human” alone does not establish compatibility with every human-antibody format.
Record amounts using the dedicated workflow
Follow this reagent’s manual to mix the test antibody with the labeling reagent and prepare the cell-assay working solution. Across antibodies, keep final antibody concentration, reagent ratio and addition volume consistent, and check final glycerol and other formulation components. The mixing ratio is not a covalent labeling density and does not imply the input or purification rules used for TFP or maleimide dyes.
Separate reagent background from antibody background
Keep medium, addition volume and acquisition conditions consistent across controls. Include an appropriate target-negative sample when assessing target dependence.
| Sample | What it checks | Compare with |
|---|---|---|
| Cells + medium | Cellular and medium background. | The baseline for all labeled conditions. |
| Cells + labeling reagent | Reagent-associated signal without the test antibody. | The complete antibody–reagent mixture. |
| Matched isotype + labeling reagent | Nontarget and Fc-associated background. | The test antibody at matched concentration and ratio. |
| Independent binding assay with unlabeled antibody | Whether labeling changes antibody binding. | Post-labeling binding performance. |
Carry screening results into follow-up validation
Record a time course and cell health for candidate antibodies. If moving to direct TFP or maleimide labeling, reassess binding and assay readout instead of pooling raw fluorescence across the two formats. In Fc-receptor-positive immune cells, evaluate target binding and Fc-associated uptake separately.
Related products and documentation
- pH630 Deep Red anti-human labeling reagent · QH-17-1003 · Request documentation
Further reading
- pHrodo selection for antibody internalization
- Controls and readouts for pHrodo internalization assays
- Troubleshooting pHrodo background and weak signal