Name
XFD750 Conjugation Kit (BSA compatible) (MAK-0290)
Cat
MAK-0290
Target
Antibody; Protein
Ex/Em (nm)
752/776
Product Type
Labeling Kit
Features
XFD750 Conjugation Kit (BSA compatible) (MAK-0290) offers several standout features:
- Rapid Conjugation: achieve XFD750 labeling in under 2 hours with minimal hands-on time.
- Compatible with antibodies containing 0.1% - 0.5% BSA.
- Labels up to 100 µg antibody.
- Versatility: suitable for conjugating antibodies and proteins. XFD750-labeled antibodies can be used immediately in applications such as Flow cytometry ICC/IF and other fluorescence imaging assays without further purification.
- Rapid Conjugation: achieve XFD750 labeling in under 2 hours with minimal hands-on time.
- Compatible with antibodies containing 0.1% - 0.5% BSA.
- Labels up to 100 µg antibody.
- Versatility: suitable for conjugating antibodies and proteins. XFD750-labeled antibodies can be used immediately in applications such as Flow cytometry ICC/IF and other fluorescence imaging assays without further purification.
Background
XFD750 has an excitation peak at 752 nm and an emission peak at 776 nm and is similar in size, brightness and application to AFluor 750, DyLight 750 and CF 750.
Product Information
The Conjugation Kit (Fast) uses a simple and quick process for Fluorescein labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. To conjugate an antibody to fluorescein using this kit:
- add modifier to antibody and incubate for 60 mins
- Purify the conjugation product through a desalting column
The conjugated antibody can be used immediately in WB, ELISA, IHC, FACS, IF etc. No further purification is required and 100% of the antibody is recovered for use. Use the to learn more about the technology, or about conjugating other proteins and peptides to Fluorescein. Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
- add modifier to antibody and incubate for 60 mins
- Purify the conjugation product through a desalting column
The conjugated antibody can be used immediately in WB, ELISA, IHC, FACS, IF etc. No further purification is required and 100% of the antibody is recovered for use. Use the to learn more about the technology, or about conjugating other proteins and peptides to Fluorescein. Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
Size
Size: 1mg
Preactivated Fluorescein: 1 vial (Protect from light)
Buffer: 50 ul
DMSO: 150 ul
Purification Column+Tube: 2 vial
Preactivated Fluorescein: 1 vial (Protect from light)
Buffer: 50 ul
DMSO: 150 ul
Purification Column+Tube: 2 vial
Recommended Ratio
1. Using the maximum amount of antibody may result in less labelling per antibody.
2. Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2mg/ml or < 0.5 mg/ml should be diluted /concentrated.
2. Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Req.
Buffer should be pH 6.5-8.5.
Sample buffer
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.
(1) Compatible buffer constituents: 50mM / 0.6% Tris(Tris buffered saline is almost always ≤ 50 mM / 0.6%), 0.1% BSA, 50% glycerol, 0.1% sodium azide ( Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.), PBS.
(2) Potassium phosphate, Sodium chloride, HEPES, Sucrose, Sodium citrate, EDTA, Trehalose.
(3) Incompatible buffer constituents: Thiomerosal, Proclin, Glycine, Arginine, Glutathione, DTT.
(4) If a constituent of the buffer containing your antibody or protein is not listed above, please contact us.
(5) Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
(1) Compatible buffer constituents: 50mM / 0.6% Tris(Tris buffered saline is almost always ≤ 50 mM / 0.6%), 0.1% BSA, 50% glycerol, 0.1% sodium azide ( Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.), PBS.
(2) Potassium phosphate, Sodium chloride, HEPES, Sucrose, Sodium citrate, EDTA, Trehalose.
(3) Incompatible buffer constituents: Thiomerosal, Proclin, Glycine, Arginine, Glutathione, DTT.
(4) If a constituent of the buffer containing your antibody or protein is not listed above, please contact us.
(5) Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Storage
Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation. -20°C
Procedure
(1) Take out the FLUORESCEIN and centrifuge for a few seconds to allow the dry powder of FLUORESCEIN in the tube to settle to the bottom;
(2) Add 100 ul of DMSO to the tube and mix repeatedly with a pipette or vortex until the FLUORESCEIN is completely dissolved;
(3) Add an appropriate amount of adjustment buffer to the antibody (10 ul of adjustment buffer per 100 ul of antibody);
(4) Add the dissolved FLUORESCEIN to the antibody (2.0-4.0 ul of FLUORESCEIN per 100 ug of antibody) and mix repeatedly with a pipette or vortex.
(5) Place the antibody-FLUORESCEIN mixture on a horizontal shaker or rotary mixer, and incubate it in the dark at room temperature (the reaction tube can be wrapped in aluminum foil) for 1 hour while shaking. Note: If a higher F/P ratio is required, the coupling time between antibody and FLUORESCEIN can be appropriately extended.
(6) Remove the purification column and centrifuge it at 3000 rpm for 2 minutes.
(7) Temporarily retain the buffer solution in the collection tube.
(8) Transfer the purification column to a new collection tube and aspirate the antibody-FLUORESCEIN conjugate onto the surface of the packing material in the purification column. If the sample volume is less than 50ul, first use 3.2 of the reserved buffer to make up the liquid volume to 50ul. The maximum sample volume should not exceed 100ul.
(9) After the sample has penetrated into the filler, centrifuge at 3000rpm for 2 minutes.
(10) Store the antibody-FLUORESCEIN conjugate in the dark at 4℃ for future use. The final product can be stored in the dark at 4℃ for up to 1 year.
(2) Add 100 ul of DMSO to the tube and mix repeatedly with a pipette or vortex until the FLUORESCEIN is completely dissolved;
(3) Add an appropriate amount of adjustment buffer to the antibody (10 ul of adjustment buffer per 100 ul of antibody);
(4) Add the dissolved FLUORESCEIN to the antibody (2.0-4.0 ul of FLUORESCEIN per 100 ug of antibody) and mix repeatedly with a pipette or vortex.
(5) Place the antibody-FLUORESCEIN mixture on a horizontal shaker or rotary mixer, and incubate it in the dark at room temperature (the reaction tube can be wrapped in aluminum foil) for 1 hour while shaking. Note: If a higher F/P ratio is required, the coupling time between antibody and FLUORESCEIN can be appropriately extended.
(6) Remove the purification column and centrifuge it at 3000 rpm for 2 minutes.
(7) Temporarily retain the buffer solution in the collection tube.
(8) Transfer the purification column to a new collection tube and aspirate the antibody-FLUORESCEIN conjugate onto the surface of the packing material in the purification column. If the sample volume is less than 50ul, first use 3.2 of the reserved buffer to make up the liquid volume to 50ul. The maximum sample volume should not exceed 100ul.
(9) After the sample has penetrated into the filler, centrifuge at 3000rpm for 2 minutes.
(10) Store the antibody-FLUORESCEIN conjugate in the dark at 4℃ for future use. The final product can be stored in the dark at 4℃ for up to 1 year.
Shipped
Ambient - Cannot Ship with Ice
Coupled Product Storage
Generally, conjugated antibodies can be stored at 4°C for up to 18 months. If longer storage is required, a cryoprotectant, such as 50% glycerol, can be used to store the conjugated antibody at -20°C. The optimal storage conditions for conjugated antibodies must be determined based on the experiment, antibody stability, and optimal storage conditions. Conjugated antibodies should be stored away from light.
