Catalog No.	PA007702.m1
Product Name	Anti-Fluorescein Monoclonal Antibody (Clone: 4-4-20 (enhanced)), | PA007702.m1
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Synonyms	Anti-FITC Antibody, Anti-fluorescein Isothyocianate Antibody
Clone	Clone: 4-4-20 (enhanced)
Isotype	Mouse IgG1 Kappa
Source/Host	The recombinant anti-Fluorescein monoclonal antibody (clone: 4-4-20 (enhanced)) was produced in mammalian cells.
Specificity/Sensitivity	The recombinant anti-Fluorescein monoclonal antibody (clone: 4-4-20 (enhanced)) specifically binds to Fluorescein.
Applications	Western Blot, Flow Cytometry.
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-fluorescein monoclonal antibody of clone 4-4-20 (enhanced) 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.
Note	Recombinant mouse IgG1 isotype controls are available. Condition of sample preparation and optimal sample dilution should be determined experimentally by the investigator.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

PA007702.m1: Recombinant Anti-Fluorescein Monoclonal Antibody (Clone: 4-4-20 (enhanced)), Mouse IgG1 Kappa

Recombinant mouse IgG1 Monoclonal Antibody.

Background of Anti-Fluorescein (FITC) Monoclonal Antibody

Recombinant mouse anti-fluorescien antibodies recognize fluorescien (FITC) attached to proteins, peptides, nucleic acids, or solid matrices, and can be offered in fluorophore and enzyme-conjugated versions.

The anti-fluorescien monoclonal antibody can be used as a second-step reagent in various applications for detecting fluorescien (FITC)-conjugated targets. The fluorescien-anti-fluorescien system is widely used in immunoassay signal amplification techniques because an anti-fluorescien antibody may bind fluorescien (FITC) with extremely high affinity.

The fluorescien-anti-fluorescien 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. Fluorescien (FITC) is not naturally present in any biological samples.

Please contact us to ask for a quote for the engineered anti-fluorescien (FITC) antibody mutants of various isotypes, and mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated antibodies and mutants. A variety of conjugates (such as dyes and fluorophores) with the anti-FITC antibodies and mutants are available.

References of Anti-Fluorescein Monoclonal Antibody:

High-affinity rat anti-fluorescein monoclonal antibody with unique fine specificity properties including differential recognition of dynamic ligand analogues.
Miklasz, S. D., et al. J Mol Recognit.1995 Jul-Aug;8(4):258-69. doi: 10.1002/jmr.300080404. PMID: 8588943
The binding properties of a monoclonal rat antifluorescein antibody specific to a given isomer were quantitatively studied to determine the capacity to bind dynamic analogues of fluorescein which exists in multiple isomers. To generate monoclonal anti-fluorescein antibodies that reacted with specific dynamic analogues of fluorescein possessing unconjugated aromatic ring systems, immune spleenocytes from Lou/M rats immunized with FITC(I)-KLH were fused with Balb/c SP2/0-Ag14 murine myeloma cells forming rat-mouse hybridomas. Thus, antibody reactivity of an anti-fluorescein antibody with phenol red and phenolphthalein was dictated more by ligand dynamics and aromatic orientation than by chemical structure similarities. P2A12-1-C8 exhibited significant reactivity with HPF and phenol red, which are dynamic structural analogues of the homologous fluorescein ligand. No reactivity was demonstrated with phenolphthalein, which based on relative chemical structures was expected to be more reactive than phenol red.
Tags: activity of Anti-Fluorescein Monoclonal; Anti-Fluorescein Monoclonal

Antibody networks and imaging: elicitation of anti-fluorescein antibodies in response to the metatypic state of fluorescein-specific monoclonal antibodies – PubMed
Cedergren, A. M., et al. Mol Immunol.1996 May-Jun;33(7-8):711-23. doi: 10.1016/0161-5890(96)00003-x. PMID: 8760283
Affinity-labeled liganded murine monoclonal anti-fluorescein antibodies served as immunogens administered both in the syngenic and xenogenic modes to determine if the metatypic state elicited anti-hapten antibodies through imaging-like mechanisms. Polyclonal and monoclonal anti-Ab1 reagents in various hosts were assayed for anti-fluorescein and/or anti-metatype specificity. Significant anti-fluorescein responses were measured indicating that the metatypic state directly or indirectly stimulates an anti-hapten antibody population. Studies are described regarding generation of anti-hapten antibodies starting with a monoclonal Ig immunogen in the ligand-induced conformation or metatypic state. Liganded monoclonal Ab1 antibodies represent the unique feature of the study since previous reports investigating internal imaging in the original Idiotype Network Hypothesis [Jerne, 1974 (Ann. Immun. 125C, 373-389)] were based on the non-liganded or idiotypic state [as reviewed in: Rodkey, 1980 (Microbiol. Rev. 44, 631-659); Kohler et al., 1979 (In: Methods in Enzymology: Antibodies, Antigens and Molecular Mimicry, pp. 3-35); Greenspan and Bona, 1993 (FASEB J. 7,437-444)].
Tags: Anti-Fluorescein Monoclonal antibody for animal model; Anti-Fluorescein Monoclonal antibody for cancer research

Pressure-induced dissociation of fluorescein from the anti-fluorescein single-chain antibody 4-4-20.
Coelho-Sampaio, T., et al. Biochemistry.1993 Oct 19;32(41):10929-35. doi: 10.1021/bi00092a001. PMID: 8218158
Hydrostatic pressure was used to dissociate fluorescein (Fl) from the high-affinity anti-Fl single-chain antibody 4-4-20 (SCA 4-4-20). This difference is explained in terms of a higher overall flexibility of unliganded SCA and of a less stable binding site in SCA relative to mAb. Fl dissociation was monitored by measuring (1) the shift in the Fl absorption peak, (2) the recovery in Fl fluorescence intensity, which is quenched upon SCA binding, or (3) the decrease in Fl fluorescence polarization. Pressure effects were studied at two different Fl:SCA 4-4-20 molar ratios: 1:1, at which Fl fluorescence quenching was ca. The dependence of KD, and consequently of delta G of dissociation, on pressure permitted calculation of the magnitude of the standard volume change (delta V) involved in the dissociation process.
Tags: Anti-Fluorescein Monoclonal antibody for mouse tumor model; Anti-Fluorescein Monoclonal for animal model

Preparation of Na,K-ATPase specifically modified on the anti-fluorescein antibody-inaccessible site by fluorescein 5′-isothiocyanate – PubMed
Lin, S. H., et al. Anal Biochem.2000 Dec 15;287(2):303-12. doi: 10.1006/abio.2000.4828. PMID: 11112278
Removal of nonspecifically bound fluorescein was also confirmed by titration of the modified Na, K-ATPase with anti-fluorescein antibody and by time resolution of the fluorescence change when the modified enzyme was mixed with Na(+) in a stopped-flow instrument. The only fluorescence change when specifically modified Na,K-ATPase was mixed with Na(+) was the signal from fluorescein at the antibody-inaccessible, substrate-protectable site that reports the conformational change in unphosphorylated enzyme. The method should be generally applicable to the preparation of specifically labeled P-type pumps for use in kinetic and equilibrium titrations or energy transfer measurements. Specific labeling is required for energy transfer measurements and to avoid artifacts in the use of fluorophores as reporter groups. Sulfhydryl reagents protected against modification of cysteine residues, and treatment with dithiothreitol eliminated a slow doubling of the fluorescence of conventionally modified Na,K-ATPase upon dilution that is attributed to disappearance of self-energy transfer.
Tags: Anti-Fluorescein Monoclonal for cancer research; Anti-Fluorescein Monoclonal for mouse tumor model

Role of electrostatic interactions in the binding of fluorescein by anti-fluorescein antibody 4-4-20.
Omelyanenko, V. G., et al. Biochemistry.1993 Oct 5;32(39):10423-9. doi: 10.1021/bi00090a018. PMID: 8399186
Anti-fluorescein antibodies are excellent model systems for studying the biochemical basis of molecular recognition because a prodigious amount of both physico-chemical and structural information is available for these antibodies. In previous studies, we determined the three-dimensional structure of an antigen-binding fragment of a high-affinity anti-fluorescein antibody (4-4-20) in complex with fluorescein. These studies showed that fluorescein binds tightly in an aromatic slot and participates in a network of electrostatic interactions. Furthermore, recombinant single-chain antibodies have been produced for several anti-fluorescein antibodies, and site-specific mutagenesis studies have defined the energetic contributions of a number of key active-site residues. The fluorescence lifetime of the 4-4-20/fluorescein complex was determined at both pH 8 and pH 10.6.
Tags: Anti-Fluorescein Monoclonal mAb; function of Anti-Fluorescein Monoclonal antibody

For more references about Anti-Fluorescein Monoclonal Antibody please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-Biotin antibodies:

In Vivo Grade Recombinant Anti-Biotin Antibodies (Clone F1)

Syd Labs provides the following recombinant anti-Fluorescein/Digoxin/Biotin/Phospho-Tyrosine monoclonal antibodies:

Recombinant Anti-Fluorescein Monoclonal Antibodies (Clone 4-4-20 (enhanced))
Recombinant Anti-Fluorescein Monoclonal Antibodies (Clone 4M5.3)
Recombinant Anti-Digoxin Monoclonal Antibodies (Clone 5A7)
Recombinant Anti-Digoxin Monoclonal Antibodies (Clone 26-10)
Recombinant Anti-Biotin Monoclonal Antibodies (Clone F1)
Recombinant Anti-Biotin Monoclonal Antibodies (Clone M33)
Recombinant Anti-Phospho-Tyrosine Monoclonal Antibodies (Clone Py20)
Recombinant Anti-Phospho-Tyrosine Monoclonal Antibodies (Clone 4G10)

Recombinant Anti-Fluorescein Monoclonal Antibody (Clone: 4-4-20 (enhanced)), Mouse IgG1 Kappa from: Recombinant Anti-Fluorescein Monoclonal Antibody (Clone: 4-4-20 (enhanced)), Mouse IgG1 Kappa: PA007702.m1 Syd Labs

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