Overview

  • Product name

    Rabbit Anti-Human IgG H&L (HRP)
    See all IgG secondary antibodies
  • Host species

    Rabbit
  • Target species

    Human
  • Tested applications

    Suitable for: Dot blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WB, ICC/IF, Competitive ELISAmore details
  • Immunogen

    human IgG whole molecule

  • Conjugation

    HRP

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...
  • Purity

    Affinity purified
  • Purification notes

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using human IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • Conjugation notes

    Horseradish Peroxidase (HRP)
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6759 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
Dot blot Use at an assay dependent dilution.
ELISA 1/130000.
IHC-P Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.
Immunomicroscopy Use at an assay dependent dilution.
WB 1/5000 - 1/25000.
ICC/IF 1/1000 - 1/5000.
Competitive ELISA Use at an assay dependent dilution. PubMed: 20573830

Images

  • ELISA results of ab6759 tested against purified human IgG protein.

    Each well was coated in duplicate with 1.0 µg of human IgG. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log 10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gelatin as blocking buffer and TMB substrate.

References

This product has been referenced in:

See all 33 Publications for this product

Customer reviews and Q&As

Application
ELISA
Cells were seeded on 96-well plates with a density of 2x105 cell/ml and grown until confluent. After 4 hours of incubation with human recombinant α-galactosidase A (AGAL, 4.6 μg/ml) and patients’ serum (50 µl per 4.6 μg/ml AGAL), cells were washed twice with PBS and lysed with passive lysis buffer (Promega, Mannheim Germany). Lysates were centrifuged (14.000x g) for 20 minutes at 4°C and supernatants were used for ELISAs, as follows. 96 well plates were coated with a capture antibody (α-AGAL, R&D Sytems, Minneapolis, US no. AF6146, 200 ng per well) over night at 4°C. After 5 washes with 200 μl PBS/Tween 0.1% and blocking with PBS/BSA 2% for 2 hours at RT, 3-10 μl of cellular lysates were applied in 100 μl PBS overnight at 4°C. Wells were washed 3 times with PBS/Tween 0.1%. For human IgG detection shuttled into cells by AGAL, anti-hIgG HRP-coupled antibodies (Abcam no: ab6759, working concentration: 0.66 μg/ml) were applied in PBS/BSA 2% for 2 hours at RT. After 5 additional washes, 100 μl 1-Step Ultra TMB-ELISA Substrate Solution (Thermo Fisher Scientific) was added to each well, incubated at RT, and stopped with 100 μl 2 mol/l sulfuric acid. Absorption was measured at 450 nm and normalized for protein content and against serum from a healthy control (negativ for antibodies against AGAL).

Figure legend: *p<0.05, **p<0.01.

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Verified customer

Submitted Sep 04 2019

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