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12158167001

Roche

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

from rat IgG1

Quality Level

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

shipped in

wet ice

storage temp.

2-8°C

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.

Specificity

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).

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.

Quality

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

Preparation Note

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

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.

Reconstitution

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

Other Notes

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

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Aquatic Chronic 3 - Skin Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 2

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

Quotes and Ordering

Edward J Hsieh et al.
Archives of biochemistry and biophysics, 463(1), 19-26 (2007-03-30)
Coenzyme Q (Q) is a redox active lipid that is an essential component of the electron transport chain. Here, we show that steady state levels of Coq3, Coq4, Coq6, Coq7 and Coq9 polypeptides in yeast mitochondria are dependent on the
Derek C Prosser et al.
Journal of cell science, 128(22), 4220-4234 (2015-10-16)
Clathrin-mediated endocytosis (CME) is a well-studied mechanism to internalize plasma membrane proteins; however, to endocytose such cargo, most eukaryotic cells also use alternative clathrin-independent endocytic (CIE) pathways, which are less well characterized. The budding yeast Saccharomyces cerevisiae, a widely used
Paul A Rowley et al.
PLoS pathogens, 12(10), e1005890-e1005890 (2016-10-07)
In eukaryotes, the degradation of cellular mRNAs is accomplished by Xrn1 and the cytoplasmic exosome. Because viral RNAs often lack canonical caps or poly-A tails, they can also be vulnerable to degradation by these host exonucleases. Yeast lack sophisticated mechanisms
Kenji Tamura et al.
Oncology letters, 14(6), 6650-6658 (2018-01-19)
The present study aimed at identifying novel molecular cancer drug targets and biomarkers by analyzing the gene expression profiles of high-grade prostate cancer (PC), using a cDNA microarray combined with laser microbeam microdissection. A number of genes were identified that
Matthew D Marsden et al.
PLoS pathogens, 13(9), e1006575-e1006575 (2017-09-22)
The ability of HIV to establish a long-lived latent infection within resting CD4+ T cells leads to persistence and episodic resupply of the virus in patients treated with antiretroviral therapy (ART), thereby preventing eradication of the disease. Protein kinase C

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