Overview

  • Product name

    Anti-Polyethylene glycol antibody [PEG-2-128]
    See all Polyethylene glycol primary antibodies
  • Description

    Rabbit monoclonal [PEG-2-128] to Polyethylene glycol
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Sandwich ELISA, WB, ELISAmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule corresponding to Polyethylene glycol conjugated to keyhole limpet haemocyanin. Synthetic peptide corresponds to Polyethylene glycol 20K (PEG20K).

  • Epitope

    This antibody recognizes the backbone of the Polyethylene glycol molecule. It can therefore be used to detect free forms of PEG. To detect conjugated forms of PEG, please see ab51257, which detects the terminal methoxy group of the PEG molecule.
  • General notes

    Antibody Isotype:  Please note, this RabMAb product's isotype is Rabbit IgM. 

    If you have any questions on our PEG products - please visit our Polyethylene glycol (PEG) FAQs page. 

    As this product's isotype is Rabbit IgM, KD values were provided by loading the antigen first before testing the antibody.
    When compared to loading the antibody first and then the antigen, this method can give approximately 10-1 difference as determined by using ab51257 - Anti-Polyethylene glycol antibody [PEG-B-47] (of rabbit IgG isotype) as a control:

    Antigen loaded first: 1.51E-10. Antibody loaded first: 2.41E-09. 

    Tween 20 usage: Tween 20 is a detergent that can interfere with ELISA and potentially other applications (often used in antibody dilution buffers, such as TBST) when using anti-PEG products. This is also the same case for other polyoxyethylene detergents.
    Therefore, it is recommended to avoid Tween 20 and other polyoxyethylene detergents when performing ELISA using anti-PEG products.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 8.24 x 10 -12 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS, Tissue culture supernatant
  • Concentration information loading...
  • Purification notes

    Harvested from hybridoma is preliminarily purified via H2O dialysis and then centrifuged to remove some impurities components.
  • Primary antibody notes

    ab133471 is useful in measuring the pharmacokinetics of Polyethylene glycol modified molecules.
  • Clonality

    Monoclonal
  • Clone number

    PEG-2-128
  • Isotype

    IgM
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab133471 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
Sandwich ELISA Use at an assay dependent concentration.
WB 1/1000. The MW of the WB band for ab133471 varies based on the PEGylated molecule detected.
ELISA Use a concentration of 2 - 10 µg/ml.

Target

  • Alternative names

    • PEG antibody

Images

  • Direct ELISA antigen dose-response curve using purified ab133471. Antigen concentration of 0-10000 ng/mL. A HRP-conjugated goat anti-rabbit IgM (1/1000) was used as the secondary antibody.

  • Direct ELISA antigen dose-response curve using purified ab133471. Antigen concentration of 0-100 ng/mL. A HRP-conjugated goat anti-rabbit IgM (1/1000) was used as the secondary antibody.

  • Sandwich ELISA antigen dose-response curve using purified ab133471.

    Capture antibody - ab133471 (10000 ng/ml).

    Antigens - MBP-PEG20K and ADI-PEG (0-1000 ng/ml).

    Detection - ab53449 (500 ng) anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) followed by peroxidase-conjugated streptravidin (1/1000).

  • All lanes : Anti-Polyethylene glycol antibody [PEG-2-128] (ab133471) at 1/1000 dilution (purified)

    Lane 1 : PEG20K
    Lane 2 : MBP-PEG20K

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgM, (H+L) at 1/500 dilution


    Blocking buffer: 15% BSA/TBS.

    Dilution buffer: 10% PAA /TBST.

  • All lanes : Anti-Polyethylene glycol antibody [PEG-2-128] (ab133471) at 1/1000 dilution (unpurified)

    Lane 1 : MBP-PEG20K at 1 µg
    Lane 2 : MBP-PEG20K at 0.5 µg
    Lane 3 : PEG20K at 1 µg
    Lane 4 : PEG20K at 0.5 µg

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgM at 1/2000 dilution


    Gel concentration: 4-20%
  • Various concentrations of Polyethylene glycol polymers with amine end groups were coated in 96 wells. 2 ug/ml of unpurified ab133471 was added. After washing, samples were incubated with 1 ug/ml of HRP-conjugated anti-rabbit IgM.

  • Various concentrations of Polyethylene glycol 20K (PEG20K) conjugated MBP and branched Polyethylene glycol conjugated BSA were coated in 96 wells. 2 ug/ml of unpurified ab133471 was added. After washing, samples were incubated with 1 ug/ml of HRP-conjugated anti-rabbit IgM.

  • 10 ug/ml of unpurified ab133471 was coated into 96 wells. Serial dilution of Polyethylene glycol 20K (PEG20K) conjugated MBP and PEGylated ADI were added. After washing, 1 ug/ml of biotinylated anti-Polyethylene glycol antibody was added. Streptavidin-HRP was then used to develop the color.

  • Method:APS sensor + Antigen (PEG 5K-BSA at 0.3µg/ml)+ antibody (ab133471 at  0.0312, 0.0625, 0.125, 0.25, 0.5, 1 µg/m).

    Sample ID: PEG-2-128

    KD (M): 8.24E-12

    kon(1/Ms): 1.47E+07

    kon Error: 3.59E+04

    kdis(1/s): 1.22E-04

    kdis Error: 6.46E-07

    Full X^2: 6.0601

    Full R^2: 0.9985

References

This product has been referenced in:

  • Sanchez MD  et al. Development and evaluation of a host-targeted antiviral that abrogates herpes simplex virus replication through modulation of arginine-associated metabolic pathways. Antiviral Res 132:13-25 (2016). WB . Read more (PubMed: 27192555) »
  • Shiraishi K  et al. Exploring the relationship between anti-PEG IgM behaviors and PEGylated nanoparticles and its significance for accelerated blood clearance. J Control Release 234:59-67 (2016). Sandwich ELISA . Read more (PubMed: 27164541) »
See all 4 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question
Answer

The antibody ab133471 cannot be used with our PEG ELISA kit ab138914.

Read More

Answer

Abcam's PEG ELISA kit ab138914 cannot be used in detecting free PEG.

Since free PEG and conjugated PEG have different MW, these two forms can be distinguished by western blot using anti-PEG (backbone) antibody, ab133471. Since ab133471 recognizes PEG backbone, PEG is not required to be bound to other molecules. We have direct ELISA data using PEG5K, 6K, 20K to confirm this.

Read More

Question
Answer

The antibody ab51257 is specific to the terminal of PEG molecule and is made up of rabbit IgG. In contrast, ab133471 is specific to the backbone of the PEG molecule and is made up of rabbit monoclonal IgM.

Read More

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