The Protein A-mediated binding of Staphylococcus to antibodies in flow cytometric assays and its reduction using FcR blocking reagent

The Protein A-mediated binding of Staphylococcus to antibodies in flow cytometric assays and its reduction using FcR blocking reagent

Staphylococcus aureus and other coagulase-positive Staphylococcus spp. bind the Fc region of IgG antibodies through the expression of protein A (SpA). This species has consequently become a source of false positive signals in antibody-based test designed to detect bacteria other targets. In this work, flow cytometry was used to study the effect of a number of factors in SpA single-cell-mediated binding to anti-human IgG antibody, including: tension; heat kills; Overnight storage; growth phase; cell physiology; surface adhesion; and growth in food model systems. Co-staining cells through antibody-stained with a dye permeability, propidium iodide and Calcein Violet AM, the physiological status of cells associated with antibody-mediated binding SpA.

Generally, cells lacking permeabilised esterase activity is not strong antibody binding. Binding of a number of commercially available polyclonal IgG antibodies to non-Staphylococcus spp. also marked. Not all species of SpA-expressing strong showing binding of mouse IgG, and one species is not known to express a strong showing SpA binding. Most SpA-expressing strain rabbit IgG antibody bound to a certain extent, whereas only one strain bound goat IgG. To reduce or eliminate the binding of IgG-mediated SpA, the following products are evaluated as a blocking reagent and applied before staining with primary or secondary antibody: normal rabbit serum, IgG isotype control, goat IgG and commercial FCR blocking reagent.

Only FCR blocking reagent consistently reduce SpA-mediated binding of Staphylococcus spp. for antibodies to other species and can be recommended as a blocking reagent in an immunoassay designed to detect non-Staphylococcus species.Importance This study characterizes a widespread problem, but little studied associated with antibody-based detection of microbes – Staphylococcus Protein A (SpA) -mediated binding of IgG antibodies – and offered a solution: the commercial use FCR blocking reagent.

A common source of false positive signals in the detection of microbes in clinical, food or environmental samples can be removed by following the findings of the authors. Using flow cytometry, the author shows the degree of heterogeneity in this culture SpA-mediated antibody binding and that the level of antibody-mediated binding SpA strain, phase- and matrix dependent growth of food and affected by simulated food processing care and adherence of cells.

In addition, studies SpA-mediated binding of Staphylococcus spp. for antibodies against other bacterial species produce highly nuanced images, leading the authors to recommend testing against multiple strains of S. aureus and S. hyicus all the antibodies to be incorporated into any immunoassay designed to detect non-Staphylococcus spp.

The Protein A-mediated binding of Staphylococcus to antibodies in flow cytometric assays and its reduction using FcR blocking reagent
The Protein A-mediated binding of Staphylococcus to antibodies in flow cytometric assays and its reduction using FcR blocking reagent

ETS transcription factor ESE-1 / Elf3 an independent prognostic factor of survival in breast cancer patients with HR + HER2 +

Objective: ETS transcription factor ESE-1 has been shown to be important in breast cancer HER2 + and ESE-1 mRNA expression has been shown to associate with the results prognosis in HER2 + subtypes, as well as in ER +, HER2 + patients with luminal B. However, the clinical significance ESE-1 protein expression is still unknown. The purpose of this current exploratory study was to evaluate the prognostic value of ESE-1 protein expression in breast cancer molecular subtypes with particular emphasis on HER2 + hormone receptor positive (HR + HER2 +) and HER2 positive (HER2 + -only) breast cancer patients.

Rabbit Anti-Goat IgG (Fc) colloidal gold conjugated (40 nm, OD10)

CG-30319 500 ul
EUR 225

Goat Anti-Rat IgG (Fc) colloidal gold conjugated (40 nm, OD10)

CG-50119 500 ul
EUR 225

Goat Anti-mouse IgG (H+L) colloidal gold conjugated (40 nm, OD10)

CG-40320 500 ul
EUR 225

Goat anti Mouse IgG (10 nm Gold Colloid)

43R-IG084GD 1 ml
EUR 705
Description: Goat anti Mouse IgG secondary antibody (10 nm Gold Colloid)

5 nm Colloidal Gold Conjugated Goat Anti-mouse IgG (H+L) secondary antibody

GA1003 1ml
EUR 172

Goat Anti-Human IgG (H+L) colloidal gold conjugated (40 nm, OD10)

CG-10319 500 ul
EUR 225

Goat anti-rabbit IgG (H+L) colloidal gold conjugated (40 nm, OD10)

CG-20319 500 ul
EUR 225

Rabbit anti-Goat IgG (H+L) colloidal gold conjugated (40 nm, OD10)

CG-30219 500 ul
EUR 225

5 nm Colloidal Gold Conjugated Goat Anti-rabbit IgG (H+L) secondary antibody

GA1013 1ml
EUR 172

5 nm Colloidal Gold Conjugated Rabbit Anti-goat IgG (H+L) secondary antibody

GA1023 1ml
EUR 172

DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

RMG-10-1064 1 mL
EUR 1022

DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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EUR 1022

DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Anti-Mouse IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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EUR 1022

Colloidal Gold (unconjugated) 10 nm, 1 OD

CG-10NM 100 ml
EUR 415

Rabbit anti-Mouse IgG (H+L) colloidal gold conjugated (40 nm, OD10

CG-40319 500 ul
EUR 225

Rabbit Anti-Human IgG (Fc) colloidal gold conjugated (40 nm, OD10)

CG-10119-R 500 ul
EUR 225

Rabbit Anti-Rat IgG (Fc) colloidal gold conjugated (40 nm, OD10)

CG-50119-R 500 ul
EUR 225

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

RRG-10-1064 1 mL
EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

RRG-10-808 1 mL
EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

RRG-10-900 1 mL
EUR 1022

DiagNano Anti-Rabbit IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

RHG-10-1064 1 mL
EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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EUR 1022

DiagNano Anti-Human IgG conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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Goat Anti-Mouse IgG (H+L)-Gold (~10 nm) Conjugate

40319-G10 0.20 ml
EUR 202

5 nm Colloidal Gold Conjugated Protein A

GA1053 1ml
EUR 172

5 nm Colloidal Gold Conjugated Protein G

GA1055 1ml
EUR 172

Streptavidin colloidal gold conjugated (40 nm, OD10)

CG-ST40 500 ul
EUR 225

Rabbit Anti-Goat IgG (H+L)-Gold (~10 nm) Conjugate

30219-G10 0.20 ml
EUR 202

Rabbit Anti-Goat IgG (H+L)-Gold (10 nm) conjugate

30222G 0.5 ml
EUR 202

Goat Anti-Human IgM (H+L) colloidal gold conjugated (40 nm, OD10)

CG-10219 500 ul
EUR 250

Goat anti Mouse IgG (20 nm Gold Colloid)

43R-IG087GD 1 ml
EUR 705
Description: Goat anti Mouse IgG secondary antibody (20 nm Gold Colloid)

Protein A colloidal gold conjugated (40 nm, OD10)

CG-PRTA40 500 ul
EUR 225

Protein G colloidal gold conjugated (40 nm, OD10)

CG-PRTG40 500 ul
EUR 225

Goat anti Rabbit IgG (15 nm Gold Colloid)

43R-IG085GD 1 ml
EUR 705
Description: Goat anti Rabbit IgG secondary antibody (15 nm Gold Colloid)

Goat anti Rabbit IgG (20 nm Gold Colloid)

43R-IG086GD 1 ml
EUR 732
Description: Goat anti Rabbit IgG secondary antibody (20 nm Gold Colloid)

Goat anti Rabbit IgG (5 nm Gold Colloid)

43R-IG100GD 1 ml
EUR 705
Description: Goat anti Rabbit IgG secondary antibody (5 nm Gold Colloid)

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

RHA-10-700 1 mL
EUR 1022

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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EUR 1022

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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EUR 1022

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

RHM-10-780 1 mL
EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

RHM-10-808 1 mL
EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

RHM-10-850 1 mL
EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

RHM-10-900 1 mL
EUR 1022

DiagNano Anti-Human IgM conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

RHM-10-980 1 mL
EUR 1022

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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EUR 1022

DiagNano Anti-Human IgA conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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Mouse monoclonal anti-Human IgG1 (Fc) colloidal gold conjugated (40 nm, OD10)

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EUR 250

Protein A/G colloidal gold conjugated (40 nm, OD10)

CG-PRTAG40 500 ul
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DiagNano Fluorophore Labeled Gold Nanoparticles, 10 nm

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DiagNano Silica-Coated Gold Nanoparticles, 10 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Galactose conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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EUR 1022

DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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DiagNano NeutrAvidin conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Streptavidin conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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EUR 1022

DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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EUR 1022

DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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EUR 1022

DiagNano GSH conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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Gold Colloid (10 nm)

62R-GC010 100 ml
EUR 467
Description: 10 nm Colloidal Gold particle suspension

Colloidal Gold (unconjugated) 20 nm, 1 OD

CG-20NM 100 ml
EUR 415

Colloidal Gold (unconjugated) 40 nm, 1 OD

CG-40NM 100 ml
EUR 354

DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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EUR 1022

DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 750 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 780 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 808 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 850 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Protein A conjugated Gold Nanorods, diameter 10 nm, absorption max 980 nm

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DiagNano Silica-Coated PEGylated Gold Nanoparticles, 10 nm

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DiagNano Gold Nanorods, diameter 10 nm, absorption max 1064 nm

BR-10-1064 25 mL
EUR 720

DiagNano Gold Nanorods, diameter 10 nm, absorption max 1200 nm

BR-10-1200 5 mL
EUR 1022

DiagNano Gold Nanorods, diameter 10 nm, absorption max 1400 nm

BR-10-1400 5 mL
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DiagNano Gold Nanorods, diameter 10 nm, absorption max 2100 nm

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DiagNano Gold Nanorods, diameter 10 nm, absorption max 600 nm

BR-10-600 25 mL
EUR 720

DiagNano Gold Nanorods, diameter 10 nm, absorption max 650 nm

BR-10-650 25 mL
EUR 720

DiagNano Gold Nanorods, diameter 10 nm, absorption max 700 nm

BR-10-700 25 mL
EUR 585

DiagNano Gold Nanorods, diameter 10 nm, absorption max 750 nm

BR-10-750 25 mL
EUR 585

DiagNano Gold Nanorods, diameter 10 nm, absorption max 780 nm

BR-10-780 25 mL
EUR 585

DiagNano Gold Nanorods, diameter 10 nm, absorption max 808 nm

BR-10-808 25 mL
EUR 585

DiagNano Gold Nanorods, diameter 10 nm, absorption max 850 nm

BR-10-850 25 mL
EUR 585

DiagNano Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Gold Nanorods, diameter 10 nm, absorption max 950 nm

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DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 1064 nm

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DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 700 nm

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DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 750 nm

OR-10-750 1mL
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DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 780 nm

OR-10-780 1 mL
EUR 1022

DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 808 nm

OR-10-808 1 mL
EUR 1022

DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 850 nm

OR-10-850 1 mL
EUR 1022

DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 900 nm

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DiagNano Organic Gold Nanorods, diameter 10 nm, absorption max 980 nm

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EUR 1022


Methods: We developed a mouse monoclonal anti-ESE-1 antibody, verified the specificity, epitope, and used immunohistochemical staining to assess ESE-1 expression in IBC approved records of 957 breast tumor samples.

Nathaniel

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