RecombinantRabMAb

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

Reviews (1)Q&A (5)References (12)
ELISA - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • Sandwich ELISA - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • Western blot - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • ELISA - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • ELISA - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • Western blot - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
  • Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Biotin Rabbit monoclonal [PEG-B-47b] to Polyethylene glycol
  • Suitable for: IHC-P, Sandwich ELISA, WB, ELISA
  • Conjugation: Biotin

Overview

  • Product name

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

  • Description

    Biotin Rabbit monoclonal [PEG-B-47b] to Polyethylene glycol

  • Host species

    Rabbit

  • Conjugation

    Biotin

  • Tested applications

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

  • Immunogen

    Chemical/ Small Molecule. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: BSA-PEG5K. IHC-P: Human cerebral cortex tissue

  • General notes

    The smallest PEG molecule which our Polyethylene glycol antibody (ab53449) 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. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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.

  • Dissociation constant (KD)

    KD = 2.41 x 10 -9 M
    Learn more about KD

  • Storage buffer

    pH: 7.2
    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

The Abpromise guarantee

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 (1)
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.
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.

Target

  • Alternative names

    • PEG antibody

Images

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

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

    Biotin conjugate (ab53449) available for this clone.

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

    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).

  • Western blot - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
    Western blot - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
    Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (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.

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

    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.

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

    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).

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

    All lanes : BSA-PEG

    Predicted band size: 2 kDa

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

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (12)

Publishing research using ab53449? Please let us know so that we can cite the reference in this datasheet.

ab53449 has been referenced in 12 publications.

  • Stock AA  et al. Conformal Coating of Stem Cell-Derived Islets for ß Cell Replacement in Type 1 Diabetes. Stem Cell Reports 14:91-104 (2020). PubMed: 31839542
  • 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). 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). PubMed: 25488654
  • der Valk FM  et al. Prednisolone-containing liposomes accumulate in human atherosclerotic macrophages upon intravenous administration. Nanomedicine N/A:N/A (2015). PubMed: 25791806
  • Jen J  et al. Pegvisomant bioavailability of single 30 mg/mL subcutaneous injection compared to two 15 mg/mL subcutaneous injections: a pharmacokinetic, safety and tolerability study. Growth Horm IGF Res 23:114-9 (2013). ELISA . PubMed: 23651793
View all Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Question

What size of the branched and linear PEG chain does this antibody react to. Will it react to atleast 500 kDa chain?

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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.

Read More

ELISA abreview for Anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin)

 Excellent
Abreviews
abreview image
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
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Abcam user community

Verified customer

Submitted Nov 20 2014

Question

Is ab53449 purified?

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Answer

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

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Question

What is the difference between ab51257 and ab53449?

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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

Which PEG antibody do you recommend in detecting linear PEG? When detecting branched PEG?

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Answer

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

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Question

I found in your catalog the following antibodies: ab73909 and ab53449, which I would like to have. Could you please tell me how much these antibodies cost, how long will it take to deliver them? Is it possible to make them without glycerol, BSA and sodium azide?
Best regards

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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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