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

Overview

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot. Store at -20°C.
  • Storage bufferpH: 7.40
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberPEG-B-47
  • IsotypeIgG
  • Research areas

Applications

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.
IHC-P 1/100.

This antibody only works on the tissues when the animals are injected with a PEGylated protein.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Alternative names
    • PEG antibody

Anti-Polyethylene glycol antibody [PEG-B-47] images

  • 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)

  • 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.

  • 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.

  • ab51257, unpurified,  staining mouse kidney at 1:10 dilution. Left panel: without animal injection with a PEGylated protein. Right panel: with animal injection with a PEGylated protein.

  • 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 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.
  • Immunoblot of different antigen-Polyethlene glycol preparations.

    Western Blot images using ab51257, unpurified, to detect BSA-PEG.

    Lane 1: 300 ng/ml;
    Lane 2: 75 ng/ml;
    Lane 3: 15 ng/ml;
    Lane 4: 1.5 ng/ml.

  • ab51257, unpurified, staining mouse muscle at 1:10 dilution. Left panel: without animal injection with a PEGylated protein. Right panel: with animal injection with a PEGylated protein.

  • ab51257, unpurified,  staining mouse liver at 1:10 dilution. Left panel: without animal injection with a PEGylated protein. Right panel: with animal injection with a PEGylated protein.

  • ab51257, unpurified, staining mouse spleen at 1:10 dilution. Left panel: without animal injection with a PEGylated protein. Right panel: with animal injection with a PEGylated protein.

  • Sandwich ELISA-Left Graph generated using unpurified ab51257 and right using purified ab51257 at 1μg/mL. Antigen concentration range 0.0156 - 1 μg/mL for PEG-IFN-α-2b. Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

  • Direct ELISA-Left Graph generated using unpurified ab51257 and right using purified ab51257 at 1μg/mL. Antigen concentration range 0.0039 - 0.25μg/mL for PEG-BSA. Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

  • Direct ELISA-Left Graph generated using unpurified ab51257 and right using purified ab51257 at 1μg/mL. Antigen concentration range 0.0156 - 1 μg/mL for ADI-PEG. Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

  • Direct ELISA-Left Graph generated using unpurified ab51257 and right using purified ab51257 at 1μg/mL. Antigen concentration range 0.0078 - 0.5 μg/mL for both PEG5000 and PEG20000. Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

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

This product has been referenced in:
  • Weber A  et al. Selective functional activity measurement of a PEGylated protein with a modification-dependent activity assay. J Pharm Biomed Anal 132:207-214 (2017). ELISA . Read more (PubMed: 27764736) »
  • Rasmussen CE  et al. Evaluation of Nonacog Beta Pegol Long-term Safety in the Immune-deficient Rowett Nude Rat (Crl:NIH-Foxn1rnu ). Toxicol Pathol N/A:N/A (2016). ICC/IF . Read more (PubMed: 26940713) »

See all 33 Publications for this product

Product Wall

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
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Abcam user community

Verified customer

Submitted Nov 08 2016

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-in...

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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.

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

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.

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


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...

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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 lin...

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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 ...

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Thank you for your enquiry regarding ab51257.

I have spoken to my colleagues in the Lab and they have confirmed me that the concentration is batch dependent but the actual concentration of this product is printed on the label of the vial. Read More

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