Product nameFITC Conjugation Kit (Fast) - Lightning-Link®
See all FITC kits
FITC Conjugation Kit / FITC Labeling Kit (Fast) (ab188285) uses a simple and quick process for FITC / Fluorescein labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.
To conjugate an antibody to FITC / Fluorescein using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 mins
The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.
Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to FITC.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid Fluorescein Labeling Kit. 310-0005 is the same as the 100 ug size. 310-0010 is the same as the 3 x 100 ug size. 310-0030 is the same as the 3 x 10 ug size. 310-0015 is the same as the 1 mg size.
Amount and volume of antibody for conjugation to FITC
Kit size Recommended
amount of antibody1
amount of antibody
3 x 10 µg 3 x 10 µg 3 x 20 µg 3 x 10 µL 100 µg 1 x 100 µg 1 x 200 µg 1 x 100µL 3 x 100 µg 3 x 100 µg 3 x 200 µg 3 x 100 µL 1 mg 1 x 1 mg 1 x 2 mg 1 x 1 mL
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 > 2 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Compatible buffer constituents
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.
50mM / 0.6% Tris1 0.1% BSA2 50% glycerol 0.1% sodium azide PBS Potassium phosphate Sodium chloride HEPES Sucrose Sodium citrate EDTA Trehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 BSA can also interfere with the use of the conjugated antibody in tissue staining.
Incompatible buffer constituents
Thiomerosal Proclin Glycine Arginine Glutathione DTT
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.
Storing and handling conjugation kits
Lyophilized Lightning-Link® components are hygroscopic.
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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg ab273993 - FITC Mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg ab273994 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab273995 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
- Antibody Concentration And Clean-Up Kit (ab102778)
- Antibody Purification Kit (Protein A) (ab102784)
- TCS Antibody Purification Kit (ab109207)
- Serum Antibody Purification Kit (Protein A) (ab109209)
- Mouse Antibody Purification Kit (ab128745)
- Mouse TCS Antibody Purification Kit (ab128749)
- BSA Removal Kit (ab173231)
Rey, Elizabeth G., Julia L. Finkelstein, and David Erickson used FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285) as part of examining folate concentration in human serum. They used the kit to conjugate FITC to FA-BSA conjugates for use in lateral flow assay.
Schematic of sample processing and lateral flow assay, and human serum results. (a) Combination of serum sample with high-pH buffer. (b) Heating of serum solution. (c) Addition of acidic buffer to lower pH. (d) Application of prepared serum solution to LFA. (e) Application of running buffer. (f) Addition of FITC conjugates to nitrocellulose membrane. Human serum results. (g) Mean T/C ratio versus folate concentration for 24 human serum samples. Error bars shown are standard deviation, n = 3. Dotted line shows four-parameter logistic curve fit. (h,i) Images of fluorescent signal from test strip for low and high folate concentration serum, respectively.
Albus, Alexandra, et al used FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285) as part of examining Naturally occurring autoantibodies (nAb)-Secreting B1 Cells. They used the kit to conjugate FITC to anti-α-Synuclein antibody for use in flow cytometry.
nAbs-secreting B1 cells are rare and highly specific. (A–F) Gating strategy to sort only CD20+ CD27+ CD43+ CD69− IgG+, and Aβ+ (E) /α-Syn+ (F) B cells. Percentages above each plot indicate the proportion of cells resulting from the previous gate. Arrows indicate single cells in gate R5. CD, cluster of differentiation; FSC, forward scatter; SSC, sideward scatter.
Baranowski, Andreas, et al used FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285) as part of examining whether the bioactive coating of calcium phosphate cements (CPCs) with bone sialoprotein (BSP) results in increased bone formation. They used the kit to conjugate FITC to bone sialoprotein prior to the coating of a three-dimensional printed CPC scaffolds. After several washing steps, the remaining BSP (green) was visualized via microscope and the BSP coating was thus qualitatively evaluated.
Robinson, Andrew P., et al used FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285) as part of characterizing oligodendroglial populations. They used the kit to conjugate FITC to Rat anti-PDGFRalpha antibody, clone APA5, for use in flow cytometry.
SJL/J mice were immunized with PLP139–151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n = 5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45− or CD45low. (B) Oligodendroglial cells were defined by double positive staining: A2B5+PDGFRα+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes.
Do, Danh C., et al used FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285) as part of examining Bla g 2 uptake by human basophils. They used the kit to conjugate FITC to cockroach allergen Bla g 2 for use in immunocytochemistry.
Human basophils were incubated with FITC conjugated Bla g 2 (FITC‐Bla g 2, green) for overnight at 37°C, washed, and analyzed by fluorescent microscopy. Nucleus was stained with the DAPI (blue).
Lydon et al. used ab188285 FITC Conjugation Kit to label mouse anti-sheep CD34 antibody with FITC.
Data shows a) Representative light-scatter (SSC-A) vs fluorescence (CD34-FITC) plot of CD34 expression on cells from ovine apheresis samples using two-colour flow cytometry. B) IgG control.
ab188285 has been referenced in 39 publications.
- Rimsa R et al. Lung on a Chip Development from Off-Stoichiometry Thiol-Ene Polymer. Micromachines (Basel) 12:N/A (2021). PubMed: 34064627
- Charras A et al. JAK Inhibitors Suppress Innate Epigenetic Reprogramming: a Promise for Patients with Sjögren's Syndrome. Clin Rev Allergy Immunol 58:182-193 (2020). PubMed: 31165348
- Xu Z et al. In Vivo Assembly of Nanoparticles Achieved through Synergy of Structure-Based Protein Engineering and Synthetic DNA Generates Enhanced Adaptive Immunity. Adv Sci (Weinh) 7:1902802 (2020). PubMed: 32328416
- Nishiyama K et al. Rapid detection of anti-H5 avian influenza virus antibody by fluorescence polarization immunoassay using a portable fluorescence polarization analyzer. Sens Actuators B Chem 316:128160 (2020). PubMed: 32322135
- Wang J et al. Suberoylanilide hydroxamic acid alleviates orthotopic liver transplantation-induced hepatic ischemia-reperfusion injury by regulating the AKT/GSK3ß/NF-?B and AKT/mTOR pathways in rat Kupffer cells. Int J Mol Med 45:1875-1887 (2020). PubMed: 32236599