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RecombinantRabMAb

Recombinant Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

Reviews (2)Q&A (9)References (50)
Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • Functional Studies - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
  • Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [PEG-B-47] to Polyethylene glycol
  • Suitable for: ELISA, WB
  • Reacts with: Species independent

Conjugates logo Related conjugates and formulations

Carrier Free HRP

Overview

  • Product name

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

  • Description

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

  • Host species

    Rabbit

  • Specificity

    This antibody recognizes the terminal methoxy group of the PEG molecule. This antibody does not cross react with non-specific targets in blood or serum. Both free form of PEG (unconjugated version) and conjugated PEG were tested by sandwich ELISA using ab51257; results showed that ab51257 only detects conjugated forms of PEG. 

  • Tested applications

    Suitable for: ELISA, WBmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

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

  • Positive control

    • IHC-P: Mouse injected with PEGylated protein kidney, muscle, liver and spleen tissue. WB: BSA-PEG.

  • General notes

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab51257 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
ELISA
Use a concentration of 0.5 - 5 µg/ml.
WB
Use a concentration of 0.015 - 0.075 µg/ml.
Notes
ELISA
Use a concentration of 0.5 - 5 µg/ml.
WB
Use a concentration of 0.015 - 0.075 µg/ml.
Application notes
Is unsuitable for IHC-P.

Target

  • Alternative names

    • PEG antibody

Images

  • Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Lane 1 : Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 1/1000 dilution (Purified)
    Lane 2 : Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 1/5000 dilution (Purified)

    All lanes : Bovine serum albumin (BSA) and PEG 5000 conjugated protein

    Lysates/proteins at 0.015 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 120 seconds


    Blocking/Diluting buffer and concentration 5% NFDM/TBST

  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

    Comparison of sandwich ELISA using RabMAb/RabMAb (ab51257/ab51257) and MAb/RabMAb (Vendor A/ab51257) for capture/detection.

    Ab51257/ab51257*: Plate coated with 5 ug/mL of #47; 5 ug/mL of #47 used for detection (*Anti-PEG 47 biotin labeled).

    Vendor A/ ab51257*: Plates coated with 100 ug/mL of Vendor A Mouse MAb; 5 ug/mL of #47 used for detection (*Anti-PEG 47 biotin labeled).

  • Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 0.015 µg/ml + BSA-PEG at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution


    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Functional Studies - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Functional Studies - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

    Method：Protein A sensor + Antigen (PEG 5K-BSA at 0.3µg/ml)+ antibody (ab51257 at  0.0312, 0.0625, 0.125, 0.25, 0.5, 1 µg/m).

    Results:

    Sample ID: PEG-5K-BSA

    KD (M): 2.41E-09

    kon(1/Ms): 2.67E+04

    kon Error: 6.81E+02

    kdis(1/s): 6.43E-05

    kdis Error: 2.92E-06

    Full X^2: 0.018444

    Full R^2: 0.982059

  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

    Comparison of 10 ug/well of activated linear (PEG5K) and branched (PEG40K) PEG using 5 ug/ml of ab51257 in Direct ELISA assay.

    Accuracy: By detecting the methoxy group of the PEG molecule itself, ab51257 is useful in measuring the pharmacokinetics of PEG-modified molecules in vivo. Data indicate that ab51257 detects various length Y-chain PEG molecules as well as single chain PEG molecules with equal affinity. ab51257 does not cross react with non-specific targets in blood or serum.

  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Comparison of ab51257 and Vendor A mouse MAb in Direct ELISA assy.

    Goat anti-rabbit IgG-AP used for anti-PEG-47 detection; goat anti-mouse IgM-AP used for Vendor A MAb detection.

    Fig 1a. Direct ELISA using 1 ug/mL of PEG-GCSF.

    Fig 1b. Direct ELISA using 1 ug/mL of PEG-IFN.

    Comparison to other anti-PEG: In both direct and sandwich ELISA assays, ab51257 shows greater affinity and accuracy than other anti-PEG antibodies when determining the concentration of PEG or PEG-modified proteins. Results were similar whether detecting PEG itself or PEG-modified targets.

  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA assay using ab51257 to detect different forms of PEG. PEG40000-BSA is a 40 kDa PEG molecule attached to BSA. PEG5000-BSA is a 5 kDa linear PEG molecule attached to BSA.
  • ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

    ELISA Graph generated using purified ab51257 at 1μg/mL.

    Antigen concentration range:

    PEG5000 and PEG20000: 0.0078 - 0.5 μg/mL
    ADI-PEG: 0.0156 - 1 μg/mL
    PEG-BSA: 0.0039 - 0.25μg/mL
    PEG-IFN-α-2b0.0156 - 1 μg/mL

    Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

  • Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)
    Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

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 (50)

Publishing research using ab51257? Please let us know so that we can cite the reference in this datasheet.

ab51257 has been referenced in 50 publications.

  • Wang Y  et al. Development of a Highly Specific Anti-drug Antibody Assay in Support of a Nanoparticle-based Therapeutic. AAPS J 22:81 (2020). PubMed: 32488626
  • Al-Mansoori L  et al. In vitro studies on CNGRC-CPG2 fusion proteins for ligand-directed enzyme prodrug therapy for targeted cancer therapy. Oncotarget 11:619-633 (2020). PubMed: 32110281
  • Seo Y  et al. Assembly and Folding Properties of Cytosolic IgG Intrabodies. Sci Rep 10:2140 (2020). PubMed: 32034177
  • Zhu S  et al. The development and characterization of a long acting anti-thrombotic von Willebrand factor (VWF) aptamer. J Thromb Haemost 18:1113-1123 (2020). PubMed: 32011054
  • Ridwan SM  et al. Distributions of intravenous injected iodine nanoparticles in orthotopic u87 human glioma xenografts over time and tumor therapy. Nanomedicine (Lond) 15:2369-2383 (2020). PubMed: 32975163
View all Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Polyethylene glycol antibody [PEG-B-47]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Universal blocking solution as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: RT°C
Fixative
Formalin
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Sharif Ridwan

Verified customer

Submitted Oct 01 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Polyethylene glycol antibody [PEG-B-47]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Macrophages)
Permeabilization
Yes - 0.3% Triton X-100
Specification
Macrophages
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 08 2016

Question

Is there a pre-established method of antigen retrieval that is known to work well for this antibody.

Read More
Answer

In our validation of ab51257 in IHC-P, antigen retrieval was not necessary. ab51257 was validated internally with mice injected intraperitoneally using 100 ug of BSA-PEG as an antigen. Heart, kidney, liver and spleen tissue were stained 4 hours post-injection using Anti-PEG RabMAb without antigen retrieval.



However, some of the papers that have used the antibody in IHC-P did use an antigen retrieval step (one example used a head mediated retrieval on a DAKO autostainer). A list of publications that used the antibody in IHC are listed below, which may be of further assistance:



http://www.ncbi.nlm.nih.gov/pubmed/24749870?dopt=Abstract

http://www.ncbi.nlm.nih.gov/pubmed/25296030?dopt=Abstract

http://www.ncbi.nlm.nih.gov/pubmed/23665258?dopt=Abstract

http://www.ncbi.nlm.nih.gov/pubmed/23788571?dopt=Abstract

http://www.ncbi.nlm.nih.gov/pubmed/22745537?dopt=Abstract

Read More

Question

What is the difference between ab51257 and ab133471?

Read More
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

Question

What is the difference between ab51257 and ab53449?

Read More
Answer

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

Read More

Question

Can the PEG ELISA kit (ab138914) be used with both linear and branched PEG?

Read More
Answer

Yes, ab51257 is used in the kit as the capture antibody in the kit and it has been shown to detect both linear and branched PEG.

Read More

Question

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

Read More
Answer

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

Read More

Question

Since this is an antibody against the methoxy group of PEG, is there a difference in its reactivity against free mPEG vs PEG coupled to a protein? Thanks!

Read More
Answer


Both free form of PEG (unconjugated version) and conjugated PEG were tested by sandwich ELISA using this anti-PEG antibody (ab51257); however, our results showed that this antibody does not detect unconjugated PEG.

If you need an antibody which detects the free form of PEG, I would recommend using an anti-PEG Backbone antibody, ab133471. This antibody recognizes PEG backbone and that PEG is not required to be bound to other molecules.

Read More

Question

Which PEG was this created with? Will it react with any PEG?

Read More
Answer

We used PEG with MW of 5K to generate ab51257. The smallest PEG molecule which our antibody can detect is PEG750. Based on customer feedback, the biggest PEG molecule detected is PEG 40K. Additionally, this product has been shown to detect both linear and branched PEG.

Read More

Question

Can you please let me know if your Epitomics anti-PEG antibodies can be used for immunohistochemistry with formalin-fixed tissues?

Read More
Answer

Thank you for contacting us and your interest in our products.

The Anti-Polyethylene glycol antibody [PEG-B-47] (catalogue number ab51257, 2061-1) has been used in IHC and is expected to be able to detect PEG if present in the tissue with IHC of formalin fixed, paraffin embedded sections.

This was performed at Epitomics with mice injected intraperitoneally using 100 ug of BSA-PEG as an antigen. Heart, kidney, liver and spleen tissue were stained 4 hours post-injection using Anti-PEG RabMAb (Cat #2061-1). Attached please find images of the results obtained.

I hope this information has been helpful. If I can be of any further assistance, please do not hesitate to let me know.

Read More

1-10 of 11 Abreviews or Q&A

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