Recombinant Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) (ab53449)


  • Product name

    Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin)
    See all Polyethylene glycol primary antibodies
  • Description

    Rabbit monoclonal [PEG-B-47b] to Polyethylene glycol (Biotin)
  • Host species

  • Conjugation

  • Tested applications

    Suitable for: IHC-P, Sandwich ELISA, WB, ELISAmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to Polyethylene glycol conjugated to keyhole limpet haemocyanin. KLH-PEG with a terminal methoxy group.

  • Positive control

    • WB: BSA-PEG5K.
  • General notes

    The smallest PEG molecule which our antibody can detect is PEG750. Based on customer feedback, the largest PEG molecule detected is PEG 40K. Additionally, ab53449 detects both linear and branched PEG based on our in-house ELISA testing. 

    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.

    Learn more about PEG RabMAb® primary antibodies and kit - PEG RabMAb Product Portal Page

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

    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.

    This product is a recombinant rabbit monoclonal antibody.


  • Form

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 2.41 x 10 -9 M
    Learn more about KD
  • Storage buffer

    pH: 7.40
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

  • Clone number

  • Isotype

  • Research areas


Our Abpromise guarantee covers the use of ab53449 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
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

For unpurified use at 1/50 - 1/100. Images of positive IHC with this monoclonal antibody, clone PEG-B-47b, are on the ab51257 datasheet.

Sandwich ELISA Use at an assay dependent concentration.
WB 1/50000. Predicted molecular weight: 2 kDa.

For unpurified use at 1/5000 - 1/20000.

ELISA Use a concentration of 1 µg/ml.

Coating plates for ELISA: 0 to 500 ng/ml.

For unpurified use at 0.5µg/ml, coating plates for ELISA: 5 to 10 ug/ml.


  • Alternative names

    • PEG antibody


  • Direct ELISA antigen dose-response curve using purified ab53449. Antigen concentration of 0-500 ng/mL. A peroxidase-conjugated steptavidin (1/1000) was used.

  • Sandwich ELISA antigen dose-response curve using purified ab53449.

    Capture antibody - Mouse anti-IFN alpha 2b (4000 ng/ml).

    Antigens - PEG-IFN alpha 2b (0-1000 ng/ml).

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

  • ELISA assay using 5 µg/mL Anti-AFP RabMAb as the capture antibody and 0.5 µg/mL Anti-PEG-47-Biotin RabMAb (unpurified ab53449) as the detection antibody. HRP-avidin (1/3000) used for anti-PEG-47 detection. This ELISA assay could detect YCA1017 peptide-PEG antigen at a concentration as low as 0.9 ng/mL.

  • Direct ELISA assay using 63 ng/mL of antigen (GCSF-PEG) to coat the plate. HRP-avidin (1/3000) was used for the detection of varying amounts of anti-PEG-47-Biotin (unpurified ab53449).

  • Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) (ab53449) at 1/50000 dilution (purified) + BSA-PEG5K at 0.02 µg

    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 2 kDa
    Observed band size: 280 kDa
    why is the actual band size different from the predicted?

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Lane 1 : Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) (ab53449) at 1/5000 dilution (unpurified)
    Lane 2 : Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) (ab53449) at 1/10000 dilution (unpurified)
    Lane 3 : Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) (ab53449) at 1/20000 dilution (unpurified)

    All lanes : BSA-PEG

    Predicted band size: 2 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue showing negative staining with purified ab53449 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.


This product has been referenced in:

  • Che L  et al. Computer-assisted engineering of programmed drug releasing multilayer nanomedicine via indomethacin-mediated ternary complex for therapy against a multidrug resistant tumor. Acta Biomater N/A:N/A (2019). Read more (PubMed: 31344512) »
  • Poirier N  et al. FR104, an antagonist anti-CD28 monovalent fab' antibody, prevents alloimmunization and allows calcineurin inhibitor minimization in nonhuman primate renal allograft. Am J Transplant 15:88-100 (2015). Read more (PubMed: 25488654) »
See all 11 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A


The smallest PEG molecule which our antibody can detect is PEG750. Based on customer feedback, the largest PEG molecule detected is PEG 40K. Additionally, ab53449 detects both linear and branched PEG based on our in-house ELISA testing.

Read More
Human Serum (Processed blood sample.)
Processed blood sample.
Sandwich (Detection)
Blocking step
BSA, Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 20 2014


Yes, ab53449 is purified before biotinylation. Protein A was used to purify this antibody.

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Both antibodies ab51257 and ab53449 are the same clone (PEG-B-47). Product ab53449 is the biotinlyated version of ab51257.

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Both ab51257 and ab53449 detect linear and branched PEG (based on our in-house ELISA results).

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Thank you for your email.

Could you let me know how many glycerol, BSA and sodium azide free vials you would like to order? Once we have this information we can then confirm the supply.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

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