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Description
pH-sensitive IgG labeling reagents plus(Green)Product Specification Molecular Weight The product has a MW of 21. 8 kDa Physical Appearance Lyophilized Powder Storage Buffer Lyophilized from sterile PBS, pH 7. 4. Normally 5 % 8% trehalose is added as protectants before lyophilization. Please see Certificate of Analysis for specific instructions of reconstitution. Reconstitution 1. Before opening the tube cap, centrifuge the sample tube at 5000g for 3 5min at room temperature to ensure the
Product Specification
| Molecular Weight | The product has a MW of 21.8 kDa |
| Physical Appearance | Lyophilized Powder |
| Storage Buffer | Lyophilized from sterile PBS, pH 7.4. Normally 5 % – 8% trehalose is added as protectants before lyophilization. Please see Certificate of Analysis for specific instructions of reconstitution. |
| Reconstitution |
1. Before opening the tube cap, centrifuge the sample tube at 5000g for 3-5min at room temperature to ensure the lyophilized sample to settie down at the bottom of the tube. 2. Dissolved lyophilized protein sample in sterile water based on the recommended volume 10uL（10ug package）. 3. After adding sterile water, cover the lid and mix them bygently tapping the tube for 5-10 times. Note: Do not vortex or vigorously pipette sample. |
| Stability & Storage |
We recommend storing the vial(s) at -20℃, desiccated and protected from light. Once reconstituted, the reagents can be stored at 2-8℃ for 1~2 weeks, or with 50% glycerol at -20℃.(Avoid repeated freezing and thawing) Store at -20'C to -80'C for 12 months in lyophilized form
|
Background
pH sensitive IgG labeling reagents provide an easy solution to measure internalization activities of antibodies. This reagent utilizes a pH-sensitive fluorescently labeled Fc binding protein that binds to IgG antibodies from various species, resulting in the formation of a fluorescently labeled antibody-reagent complex. After antibody internalization, the surrounding pH becomes acidic and significantly enhances fluorescence signal of antibody-reagent complex. The fluorescence intensity can be used as an indicator to determine the internalization activity of antibodies. By measuring the strength of the fluorescence signal, researchers can assess the efficiency of antibody internalization into cells. This information is crucial in understanding the cellular uptake mechanism of antibodies and assessing their efficacy in targeted therapies or diagnostic applications. Additionally, monitoring the fluorescence intensity can also provide insights into the kinetics of antibody internalization, helping researchers optimize experimental condition and improve the design of antibody-based drug delivery systems.
This product can becan be used for human IgG1, IgG2, IgG3, IgG4, rabbit IgG, mouse IgG1, IgG2a, IgG2b and IgG3 and can be detected with Flow cytometry FITC or AF488 filter.
Protocol
UA070080 for Detection of antibody internalization-SOP
1. UA070080 incubation with antibodies
1.1 Dissolve UA070080 in freeze-dried powder form according to datasheet using deionized water.
1.2 Mix the tested antibody with UA070080 in a mass ratio of 3.5:1 (molar ratio 1:2), and dilute the antibody with UA070080 mixture to twice the tested concentration using complete culture medium.
1.3 Incubate at room temperature in dark for 1 hour to obtain Ab-UA070080 complex 2X working solution.
2. Ab-UA070080 complex incubation with cells
2.1 Collect and wash cells, adjust cell concentration using complete culture medium.1-2×105 cells/mL for suspend cells, 0.5-1×105 cells/mL for adherent cells. Add 100μL cell suspension to each hole of the 96 well plate.
2.2 Add 100μL Ab-UA070080 complex 2X working solution to each hole. Incubate in a 5% CO2 incubator at 37℃.
3. Flow cytometry detection
After 18-24h of culture, cells were collected for flow cytometry to detect the entosis effect of antibodies using FITC or AF488 channels
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