Catalog No.	PA007522.m1
Product Name	Anti-biotin Antibody, Mouse IgG1 | PA007522.m1
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Synonyms	Biotin; Vitamin H; Vitamin B7
Clone	F1.
Isotype	Mouse IgG1 kappa.
Specificity/Sensitivity	Biotin.
Applications	ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Form Of Antibody	0.2 uM filtered solution of 1x PBS.
Endotoxin	< 1 EU per 1 mg of the protein by the LAL method.
Purity	>95% by SDS-PAGE under reducing conditions.
Shipping	The in vivo grade recombinant anti-biotin antibodies (clone of F1) 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. 1 month from date of receipt, 2 to 8°C as supplied. 12 months from date of receipt, -20 to -70°C as supplied.
Note	Recombinant anti-biotin antibodies and mutants good for in vitro and in vivo studies. The variable regions were not humanized in the human antibody versions.
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

PA007522.m1: In vivo Grade Recombinant Anti-biotin Antibody, Mouse IgG1 Kappa

The in vivo grade recombinant anti-biotin antibody, mouse IgG1 kappa was produced in mammalian cells.

Background of Recombinant Anti-biotin Antibody

Recombinant mouse anti-biotin antibodies recognize biotin (vitamin B7) attached to proteins, peptides, nucleic acids, or solid matrices, and can be offered in fluorophore and enzyme-conjugated versions.

The anti-biotin monoclonal antibody can be used as a second-step reagent in various applications for detecting biotinylated targets. The biotin-streptavidin system is widely used in immunoassay signal amplification techniques because streptavidin binds biotin with extremely high affinity.

Because biotin is naturally present in a large variety of biological samples, it may affect the results of biotin-based assays, such as reduced sensitivity, false negatives or false positives. Streptavidin/biotin blocking kit, avidin/biotin blocking system, or biotin blockers are used to block signals from endogenous avidin, biotin and biotin-binding proteins in tissues when used with biotin-based IHC detection.

Please contact us to ask for a quote for the engineered anti-biotin 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-biotin antibodies and mutants are available.

References of Anti-biotin Antibody:

1、Improved elution strategy and new monoclonal anti-biotin antibody for LC-MS/MS characterization of protein biotinylation sites.
Zhu, Y., et al. Biochem Biophys Rep. 2024 Apr 18;38:101711. doi: 10.1016/j.bbrep.2024.101711. PMID: 38681669.
“Udeshi et al. developed a method using an anti-biotin antibody to enrich biotinylated peptides, as published in Nature Methods. …The anti-biotin antibody is a good capture reagent for biotinylated peptides because of its relatively low binding affinity to biotin, which thereby ensures high recovery via increase in elution efficiency. …The approach involved developing a monoclonal anti-biotin antibody with better reproducibility as compared to existing polyclonal antibodies (pAbs). …A monoclonal anti-biotin antibody was developed at CST using a standard antibody development pipeline. …Samples were incubated with anti-biotin antibody beads for 1 h at 4 °C.”

2、Development of an ultrasensitive sandwich immunoassay for detecting small molecule semicarbazide.
Zhang, S. W., et al. Food Chem. 2023 Dec 15;429:136835. doi: 10.1016/j.foodchem.2023.136835. PMID: 37463535.
“Anti-biotin antibody-HRP and avidin-HRP were purchased from Sigma-Aldrich (St Louis, MO, USA), biotin (S)-sulfoxide and lidimycin were purchased from TopScience (Shanghai, China), and 96-well ELISA plates were purchased from Corning (USA). …The failure to develop sandwich ELISA for small molecules is not directly caused by their molecular weight but largely by the steric hindrance caused by the small distance between epitopes. …In the present study, a new derivatizing agent was prepared by coupling o-nitrobenzaldehyde and biotin with spacer arms, and two sandwich ELISA modes were established for detecting the small molecule compound SEM, namely, double-antibody sandwich and antibody-avidin sandwich. …A sandwich ELISA method was developed for SEM based on anti-SEM-NBA antibody and avidin. …Under the distances, the sensitivities of the sandwich ELISA were no lower than those of competitive ELISA.”

3、Possible frequent multiple mitochondrial DNA copies in a single nucleoid in HeLa cells.
Pavluch, V., et al. Sci Rep. 2023 Apr 8;13(1):5788. doi: 10.1038/s41598-023-33012-6. PMID: 37031254.
“Cy3B560 was conjugated with the secondary antibodies, staining anti-biotin primary antibodies, while biotin was abundantly inserted into the 7S DNA hybridization probe for mtFISH. …These are co-localized with red small spheres/points, indicating localizations of the ATP6-COX3 gene region which was target of the ATP6COX-biotin 1,100 bp hybridization probe with the directly inserted biotin, which was visualized by anti-biotin Cy3B-immunostaining. …This mtFISH probe was visualized by anti-biotin primary Ab followed by Cy3B-560-conjugates secondary antibodies (“ATP6COX-biotin-Cy3B”) (Fig. 1b). …The average photon count from a single signal detected and localized was 400. The average number of events per frame was 6.2. Imaging of Cy3B preceded imaging with A647. …Routinely, only nucleoids constructed from more than 10 localized A647 (Cy3B) fluorophores were considered in each image.”

4、Development of a novel flow cytometry method for detecting pneumococcal-specific B cells.
Tzovara, I., et al. Cytometry A. 2022 Jul;101(7):588-596. doi: 10.1002/cyto.a.24654. PMID: 35527678.
“Excess anti-biotin-PE was removed by washing the labeled bPS-beads three times. …Aggregation of the bead particles on the side of the tube allowed for easy supernatant aspiration. …The ratio of bPS-beads to anti-biotin-PE was tested in a stepwise manner at v/v ratios of: 50:0.5, 50:1, 50:2, 50:4, and 50:8 for each pneumococcal serotype. …The bead-based Ag-specific B cell quantitation method was compared with unbound bPS detected directly with anti-biotin-PE without the involvement of beads. …PBMCs were incubated with the mAbs combination described above, the bPS monomer, and anti-biotin-PE.”

5、Identification of Host Receptors for Fungi Using Whole Cell Affinity Purification.
Phan, Q. T., et al. Methods Mol Biol. 2021;2260:27-36. doi: 10.1007/978-1-0716-1182-1_3. PMID: 33405029.
“Here, we describe a method for identifying novel host cell receptors for fungi that uses intact organisms to precipitate biotin-labelled host cell membrane proteins, which are then detected by immunoblotting with an anti-biotin antibody. …Receptors on endothelial and epithelial cells often recognize molecules that are expressed by fungi, and only a limited number of these receptors have been identified to date. …Presented here is the method to use for identification of membrane proteins that bind to C. albicans.”

6、sBioSITe enables sensitive identification of the cell surface proteome through direct enrichment of biotinylated peptides.
Garapati, K., et al. Clin Proteomics. 2023 Dec 5;20(1):56. doi: 10.1186/s12014-023-09445-6. PMID: 38053024.
“In this strategy, the primary amine groups of lysines on proteins on the surface of live cells are first labeled with biotin, and subsequently, biotinylated peptides are enriched by anti-biotin antibodies and analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). …Following cell lysis and proteolytic digestion, biotinylated peptides were enriched using our previously published protocol which employs bead-immobilized anti-biotin antibodies. …Proteins were harvested and digested prior to enrichment of biotinylated peptides using anti-biotin antibodies for LC–MS/MS analysis. …We have developed sBioSITe—a novel method to enrich surface proteins by labeling them with biotin followed by enrichment at the peptide level using anti-biotin antibodies, and detection of peptides with intact biotin tags. …Our strategy combined the use of a non-cleavable biotinylation reagent with enrichment using anti-biotin antibodies, enabling us to detect peptides with biotin tags intact.”

7、Detection and Quantification of Biotinylated Sites for TurboID-Based Proximity Labeling Mass Spectrometry in Arabidopsis.
Karunadasa, S. S., et al. Methods Mol Biol. 2025;2953:127-141. doi: 10.1007/978-1-0716-4694-6_9. PMID: 40638046.
“This chapter provides a detailed, step-by-step protocol for sample preparation, including enrichment methods using both anti-biotin antibody-based and streptavidin-based affinity beads. …Additionally, it offers an overview of software tools crucial for the reliable identification and quantification of biotinylated peptides. …Proximity labeling mass spectrometry (PL-MS) is a powerful technique for mapping protein–protein interactions (PPIs), subcellular and cell type-specific proteomes, and protein–RNA and protein–DNA interactions. …Direct detection of biotinylated peptides is critical for the accurate characterization of proximity-tagged proteins. …A peptide-level enrichment workflow is advantageous as it facilitates the efficient removal of free biotin prior to enrichment, thus allowing for the use of higher concentrations of biotin—an important advantage in certain plant studies.”

8、Preliminary evaluation of a whole blood dual-platform flow cytometry protocol for CAR-T cell quantitation: Toward accreditation and clinical routine application.
Pescarmona, R., et al. J Immunol Methods. 2025 Dec;545:113986. doi: 10.1016/j.jim.2025.113986. PMID: 41135737.
“For CAR-T cell detection, we used CD19 and BCMA CAR detection reagents (i.e. biotinylated CD19 or BCMA recombinant protein) from Miltenyi Biotec (Bergisch Gladbach, Germany) in association with a phycoerythrin (PE)-conjugated anti-biotin antibody. …After a PBS wash step (6 min, 300 g) for both tubes, cells from the “CAR-T stained” condition were incubated with 0.5 μL of CD19 or BCMA CAR detection reagent for 10 min at room temperature, protected from light. …We focused on evaluating the performance characteristics of CAR-T cell proportion measurements, as the method for determining CD3+ T cell absolute counts using the AQUIOS system had already received COFRAC accreditation. …Intermediate precision refers to the analysis of the same sample under different conditions, with at least one factor varied (e.g. operator, time, reagent batches, calibrations …). …As for the repeatability, the intermediate precision of the full process was tested on Navios using 2 blood samples with low and high levels of CD19 CAR-T cells.”

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

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

In Vivo Grade Recombinant Anti-Biotin Monoclonal Antibodies (Clone F1)
In Vivo Grade Recombinant Anti-DNP Monoclonal Antibodies (Clone DNP.m1)
In Vivo Grade Recombinant Anti-Fluorescein (FITC) Monoclonal Antibodies (Clone 4-4-20 (enhanced))

In vivo Grade Recombinant Anti-biotin Antibody, Mouse IgG1 Kappa from: In vivo Grade Recombinant Anti-biotin Antibody, Mouse IgG1 Kappa: PA007522.m1 Syd Labs

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