Donkey Anti-Rabbit IgG H&L (6nm Gold) preadsorbed (ab105294)

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Overview

  • Product name

    Donkey Anti-Rabbit IgG H&L (6nm Gold) preadsorbed
    See all IgG secondary antibodies

  • Host species

    Donkey

  • Target species

    Rabbit

  • Specificity

    Recognizes the heavy chains on rabbit IgG and the light chains common to most rabbit immunoglobulins. Does not crossreact with non-immunoglobulin serum proteins. Exhibits minimal crossreactivity to bovine, chicken, goat, guinea pig, syryan hamster, horse, human, mouse, rat and sheep serum proteins.

  • Tested applications

    Suitable for: Electron Microscopy, WB, ELISA, Othermore details

  • Minimal
    cross-reactivity


    Chicken, Cow, Goat, Guinea pig, Horse, Human, Mouse, Rat, Sheep, Syrian hamster more details

  • Conjugation

    Gold 6nm

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    pH: 8.50
    Preservative: 0.05% Sodium azide
    Constituents: 0.164% Sodium phosphate, 0.381% Sodium borate, 1.5% BSA, 0.87% Sodium chloride
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab105294 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Electron Microscopy 1/20 - 1/40.
WB Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Other Use at an assay dependent concentration. Light microscopy

References

ab105294 has not yet been referenced specifically in any publications.

Publishing research using ab105294? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

Do you have an EM protocol for use of the antibody?
When are the primary and secondary antibodies added? Before or after the cells are fixed for EM?

Many thanks

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Answer

Fixation precedes incubation with primary and secondary antibodies.

The protocol for gold-antibody complexes is essentially the same as for all conjugates with the exception of the buffer used to dilute and wash.


Step 1: Block with 5% normal serum from the host species of the gold-secondary antibody diluted in PBS.

Step 2: Wash with PBS-BSA.

In each step the grid is floated on a drop of solution.



Control: Step 3: Incubate directly with the secondary antibody without addition of the primary antibody.


Step 4: Wash with PBS-BSA.

This control experiment will tell you if the secondary is giving background. If it is not giving background but there is background in the full experiment, you know that any background is coming from the primary.



Full Experiment: Step 3: Incubate with the primary antibody diluted in PBS-BSA.

Step 4: Wash with PBS-BSA.

Step 5: Incubate with the gold labeled secondary antibody.

Step 6: Wash with PBS-BSA

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Question

Konzentration? Verduennung fuer EM nicht ausrechenbar.

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Answer

Vielen Dank für Ihren Anruf.

Wie versprochen, habe ich das Labor wegen der genauen Konzentration von ab105294 kontaktiert, und habe erfahren, dass die genaue Konzentration leider nicht bestimmt werden kann. Zwar wird der Antikörper vor der Konjugation mit den Goldkomplexen aufgereinigt. Die exakte Konzentration lässt sich jedoch nicht bestimmen, weil nach der Konjugation Monomere von Multimeren über eine Dichtegradientenzentrifugation abgetrennt werden. Diese Monomere werden vor und nach der Zentrifugation mit Albumin stabilisiert, und leider gibt es keine verlässliche Methode, die Konzentration des Antikörpers in dieser Präparation zu bestimmen.

Nach unserer Erfahrung zeigt dieser sekundäre Antikörper in der EM in Verdünnungen von 1:10 bis 1:100 gute Ergebnisse, so dass wir hoffen, diese Angaben können Ihnen für Ihre Experimente als Anhaltspunkt dienen.

Ich möchte mich dafür entschuldigen, dass ich nicht mehr Informationen für Sie habe und hoffe, dass diese Angabe dennoch hilfreich für Sie ist. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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