Recombinant
RabMAb

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

Overview

  • 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
    Rabbit
  • Conjugation
    Biotin
  • 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). 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

    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.

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 or -80°C. Avoid freeze / thaw cycle.
  • 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
    Monoclonal
  • Clone number
    PEG-B-47b
  • Isotype
    IgG
  • Research areas

Applications

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.

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.

Target

  • Alternative names
    • PEG antibody

Images

  • 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).

  • 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.

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

    Secondary
    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

  • 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).

References

This product has been referenced in:
  • 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) »
  • der Valk FM  et al. Prednisolone-containing liposomes accumulate in human atherosclerotic macrophages upon intravenous administration. Nanomedicine N/A:N/A (2015). Read more (PubMed: 25791806) »
See all 10 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer



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.

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Application
ELISA
Sample
Human Serum (Processed blood sample.)
Specification
Processed blood sample.
Type
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

Question
Answer

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

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Question
Answer

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

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Question
Answer

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

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Answer

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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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