12158167001

Anti-HA-Biotin, High Affinity (3F10)

from rat IgG₁

• Synonym(s): antibody
• UNSPSC Code: 12352203
• NACRES: NA.54
• Conjugate: biotin conjugate
• Clone: 3F10, monoclonal
• Application: —
• Citations: 70

Properties

• biological source: rat
• conjugate: biotin conjugate
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: 3F10, monoclonal
• form: lyophilized (stabilized)
• packaging: pkg of 50 μg
• manufacturer/tradename: Roche
• isotype: IgG1
• epitope sequence: YPYDVPDYA
• storage temp.: 2-8°C

Description

General description

Anti-HA-Biotin, High Affinity (3F10) is a monoclonal antibody for the highly sensitive detection of HA-tagged recombinant proteins, Fab fragments, conjugated to biotin. The Anti-HA-Biotin, High Affinity antibody (clone 3F10) recognizes the same epitope as clone 12CA5. It is a monoclonal antibody whose high affinity and low working concentrations result in less cross-reactivity than with other antibodies to the HA-epitope. Anti-HA-Biotin, High Affinity (3F10) is a biotin conjugate of this clone which is specifically useful in western blotting, ELISA applications and assays using the universal biotin-streptavidin platform, by allowing specific and highly sensitive detection of HA-tagged proteins.

Immunogen

Amino acids 98-106 from the human influenza virus hemagglutinin protein

Application

Anti-HA-Biotin, High Affinity (3F10) is used for the detection of HA-tagged recombinant proteins using:

• Dot blots
• ELISA (enzyme-linked immunosorbent assay)
• Western blots

Since Anti-HA High Affinity is a rat monoclonal, it is possible to use it in conjunction with murine monoclonals for double labeling.
It has also been used for immunocytochemistry, immunofluorescence and αScreen format based assay.[1][2][3]

Biochem/physiol Actions

Anti-HA-Biotin, High Affinity (3F10) recognizes the 9-amino acid sequence YPYDVPDYA, derived from the human influenza hemagglutinin (HA) protein. This epitope is also recognized in fusion proteins regardless of its position (N-terminal, C-terminal or internal).

Preparation Note

Sample Materials
Sample preparation: Prepare protein extracts containing the HA-tagged protein of interest using any of a variety of standard methods. The following lysis buffers have performed well and should be taken as guidelines:

• Bacterial extracts: 20 mM Tris, pH 8.0, 100 mM NaCl, cOmplete Protease Inhibitor Cocktail Tablets, followed by freeze-thaw.
• Mammalian extracts: 50 mM Tris, pH 7.5, 150 mM NaCl, 0.1% Nonidet P40, complete Protease Inhibitor Cocktail Tablets.
• Other cell lysis buffers may be more appropriate for individual applications. In general, to obtain optimal performance of the affinity matrix:
• Use protease inhibitors to reduce proteolytic activity. Use complete Protease Inhibitor Cocktail Tablets for most applications.
• Limit detergent to the lowest concentration levels necessary to obtain adequate cell lysis.

Working concentration: Working concentration of conjugate depends on application and substrate

The following concentrations should be taken as a guideline:

• Dot blot: 100 ng/ml
• ELISA: 100 ng/ml
• Western blot: 100 ng/ml

Add 1 ml double-distilled water to a final concentration of 50 μg/ml.
Rehydrate for 10 minutes prior to use.

Analysis Note

Function test: The Anti-HA-Biotin; High Affinity is function tested by Western blot analysis of a HA-tagged fusion protein.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety Information

• pictograms: GHS07
• signalword: Warning
• hcodes: H317,H412
• pcodes: P261 - P273 - P280 - P333 + P313 - P362 + P364 - P501
• Hazard Classifications: Aquatic Chronic 3 - Skin Sens. 1
• Storage Class: 11 - Combustible Solids
• wgk: WGK 2
• flash_point_f: does not flash
• flash_point_c: does not flash