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Home > Antibodies > Primary Antibodies for Epitope Tags  > Anti-FITC Monoclonal Antibodies

Anti-FITC Monoclonal Antibodies

Anti-Fluorescein Antibody

Catalog No. Product Name Size List Price (US$) Quantity
PA007703.h1 Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa 200 ug 250.00
PA007703.h1 Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa 1 mg 500.00
PA007703.h1DA Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1-D265A Kappa 200 ug 250.00
PA007703.h1DA Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1-D265A Kappa 1 mg 500.00
PA007703.h1LA Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1-L234A L235A P329G (LALAPG) Kappa 200 ug 250.00
PA007703.h1LA Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1-L234A L235A P329G (LALAPG) Kappa 1 mg 500.00
PA007703.h1F-b Biotinylated Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa 200 ug 375.00
PA007703.h1F-b Biotinylated Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa 1 mg 750.00
PA007703.h1FB Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa (Flag-Avi) 200 ug 500.00
PA007703.h1FB Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa (Flag-Avi) 1 mg 1000.00
Description

Background of Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3)

Our recombinant anti-FITC antibody products recognize FITC attached to proteins, peptides, nucleic acids, or solid matrices, and can be offered in fluorophore and enzyme-conjugated versions. This high-affinity anti-FITC monoclonal antibody can be used as a second-step reagent in various applications for detecting FITC (fluorescein)-conjugated targets. The FITC-anti-FITC system is widely used in immunoassay signal amplification techniques because our anti-FITC antibody binds FITC (fluorescein) with extremely high affinity. The FITC-anti-FITC system is different from the biotin-streptavidin system because biotin is naturally present in a large variety of biological samples, and it may affect the results of biotin-based assays, such as reduced sensitivity, false negatives or false positives. In contrast, when using an anti-FITC antibody, FITC (fluorescein) is not naturally present in any biological samples. Please contact us to ask for a quote for engineered anti-FITC antibody mutants of various isotypes, mutants with Avi-, His-, and DYKDDDDK-tags, and our biotinylated anti-FITC antibody options. A variety of conjugates (such as dyes and fluorophores) with these anti-FITC monoclonal antibody variants are available.

PA007703.h1: Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3), Human IgG1 Kappa

Recombinant human IgG1 monoclonal antibody.
Clone: 4M5.3.
Isotype: Human IgG1 Kappa.
Source: The recombinant anti-FITC antibody (clone name: 4M5.3) was produced in mammalian cells.
Specificity/Sensitivity: The recombinant anti-FITC monoclonal antibody (clone: 4M5.3) specifically binds to FITC.
Applications: ELISA, Western Blot (WB), Flow Cytometry (FC).
Form of Antibody: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.

Shipping: The recombinant anti-FITC antibody of clone 4M5.3 is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70°C as supplied.
1 month from date of receipt, 2 to 8°C as supplied.

References of Recombinant Anti-FITC Monoclonal Antibody (Clone: 4M5.3)

Key Application: Analyzing femtomolar-affinity antigen-antibody interactions and structural flexibility for advanced antibody engineering.

Antigen-antibody interactions and structural flexibility of a femtomolar-affinity antibody

Biochemistry. ;51(12):2597-2605. doi: 10.1021/bi3000319. PMID: 22390639

The femtomolar-affinity mutant antibody (4M5.3) generated by directed evolution is interesting because of the potential of antibody engineering. In this study, the mutant and its wild type (4-4-20) were compared in terms of antigen-antibody interactions and structural flexibility to elucidate the effects of directed evolution. The pulling forces of SMD simulations elucidated the regions that form strong attractive interactions in the binding pocket. These landscapes were regarded as funnel-like binding free energy landscapes. It was shown that the fluctuations in the positions of the antigen and residues in contact with antigen tend to be smaller in the mutant than in the wild type.

(Tags: High-affinity Anti-FITC Antibody; Antibody Engineering; Clone 4M5.3 Activity)
Key Application: Recombinant expression, rapid refolding, and high-efficiency purification of anti-fluorescein single-chain antibodies (scFv).

Rapid refolding and polishing of single-chain antibodies from Escherichia coli inclusion bodies

Protein Expr Purif. ;26(2):301-8. doi: 10.1016/s1046-5928(02)00538-7. PMID: 12406685

Two anti-fluorescein single-chain antibodies, 4-4-20 and 4M5.3, were expressed as inclusion bodies in E. coli for use in a comparative refolding study. Although all three methods produced active protein for both 4-4-20 and 4M5.3, the extent of aggregation differed greatly among the methods. Genes for the anti-fluorescein single-chain antibodies 4-4-20 and 4M5.3 in pRS316 [17] were subcloned into pET-21a (Novagen) using the NheI and XhoI restriction sites. A significant decrease in the fluorescent intensity of a fluorescein solution is observed upon the binding of fluorescein to the active site of 4-4-20 and 4M5.3. The anti-fluorescein single-chain antibodies 4-4-20 and 4M5.3 were each expressed as inclusion bodies in E. coli as described in the Materials and methods section.

(Tags: Anti-FITC Antibody Expression; Recombinant Antibody Purification; Single-Chain Fv (scFv))

Frequently Asked Questions (FAQ) about Anti-FITC Antibody (Clone: 4M5.3)

Q: Does this Recombinant Anti-FITC Antibody cross-react with other fluorophores like Alexa Fluor 488?

No. Our recombinant anti-FITC antibody (Clone 4M5.3) is highly specific to fluorescein isothiocyanate (FITC) and free fluorescein. It shows negligible cross-reactivity with other structurally distinct green fluorophores, such as Alexa Fluor 488 or GFP, ensuring high signal specificity in multiplexing assays.

Q: What are the advantages of using Anti-FITC Antibody Clone 4M5.3 over traditional clones?

Clone 4M5.3 is a globally recognized, directed-evolution mutant antibody known for its ultra-high, femtomolar-affinity binding capabilities. Compared to the wild-type 4-4-20 clone, Clone 4M5.3 exhibits much higher structural flexibility and significantly tighter binding pocket interactions, making it perfect for sensitive immunoassay signal amplification.

Q: Can this anti-FITC monoclonal antibody be used for Flow Cytometry (FC) and Western Blot (WB)?

Yes. This recombinant anti-FITC monoclonal antibody is fully validated for ELISA, Western Blot, and Flow Cytometry. It is highly effective as a second-step detection reagent for recovering or amplifying weak signals from FITC-conjugated primary antibodies, nucleic acids, or peptides.

Q: What is the expression source and purity of your Recombinant Anti-FITC Antibody?

Our recombinant anti-FITC antibody is expressed in mammalian cells (not E. coli inclusion bodies), ensuring proper folding, post-translational modifications, and low aggregation. The final product achieves >95% purity verified by both SDS-PAGE and HPLC, providing superior batch-to-batch consistency for reproducible research.

Q: Do you offer custom formats, specific isotypes, or tags for the anti-FITC antibody?

Yes. We provide extensive custom engineering services for the anti-FITC antibody mutants. You can request various heavy/light chain isotypes, engineered Fc regions (such as D265A or LALAPG), and specific affinity tags including Avi-tag, His-tag, or DYKDDDDK-tag. Biotinylated anti-FITC antibody options are also available upon request.

Syd Labs provides the following recombinant anti-Biotin/Digoxin/DNP/Fluorescein monoclonal antibodies:
Recombinant Anti-Biotin Monoclonal Antibodies (Clone F1)
Recombinant Anti-Biotin Monoclonal Antibodies (Clone M33)
Recombinant Anti-Digoxin Monoclonal Antibodies (Clone 5A7)
Recombinant Anti-Digoxin Monoclonal Antibodies (Clone 26-10)
Recombinant Anti-DNP Monoclonal Antibodies (Clone DNP.m3)
Recombinant Anti-Fluorescein Monoclonal Antibodies (Clone 4-4-20)
Recombinant Anti-Fluorescein Monoclonal Antibodies (Clone 4M5.3)

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