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, IHC-P
- Reacts with: Species independent
Product nameAnti-Polyethylene glycol antibody [PEG-B-47]
See all Polyethylene glycol primary antibodies
DescriptionRabbit monoclonal [PEG-B-47] to Polyethylene glycol
SpecificityThis 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. To detect free form of PEG, we recommend using ab133471. ab133471 recognizes PEG backbone and that PEG is not required to be bound to other molecules.
Tested applicationsSuitable for: ELISA, WB, IHC-Pmore details
Species reactivityReacts with: Species independent
Chemical/ Small Molecule corresponding to Polyethylene glycol conjugated to keyhole limpet haemocyanin. KLH-Polyethylene glycol with a terminal methoxy group.
- IHC-P: Mouse injected with PEGylated protein kidney, muscle, liver and spleen tissue. WB: BSA-PEG.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Upon delivery aliquot. Store at -20°C.
Dissociation constant (KD)KD = 2.41 x 10 -9 M Learn more about KD
Storage bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
PurityProtein A purified
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.
|ELISA||Use a concentration of 0.5 - 5 µg/ml.|
|WB||Use a concentration of 0.015 - 0.075 µg/ml.|
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.
- PEG antibody
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.
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
Paraffin-embedded mouse kidney tissue stained for Polyethylene glycol using purified ab51257 at 1/50 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer) secondary. Counterstained with Hematoxylin. No staining on mouse kidney without PEGylated protein treatment (A), positive staining on mouse kidney with PEGylated protein treatment (B).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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).
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.
Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 0.015 µg/ml + BSA-PEG at 10 µg
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 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.
Immunohistochemistry images of anti-PEG (brown) and hematoxylin counterstaining (blue) of tumor (A1–A3 and B1–B3) and muscle tissue (A4 and B4) 30 minutes (A) and 2 h (B) post injection of Spago Pix
Tumor tissue at both time points is shown at three different enlargements where the enlarged areas are indicated by the square boxes. Vessels (V), fibroblasts (Fb) and fat cells (Fc) are indicated with rounded lines. Note that the tumor tissue in panel B1–B3 originates from a mouse injected with a dose of 10 µmol Mn/kg, whereas the other tissues originate from animals administered 20 µmol Mn/kg.
Paraformaldehyde fixed muscle and tumor tissues collected before injection and at 30 minutes and 2–4 h post-dose were paraffin embedded and thereafter cut (4 µm) and positioned on glass tissue slides. Prior to staining, the slides were placed in an antigen-retrieval solution using an automated pre-treatment module (PT-Link; Dako, Glostrup, Denmark). Slides were stained for PEG in an automated immunohistochemistry robot (Autostainer; Dako). PEG is detected with ab51257 at 0.6 μg/ml.
(From Figure 6 Eriksson et al)
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).
Sample ID: PEG-5K-BSA
KD (M): 2.41E-09
kon Error: 6.81E+02
kdis Error: 2.92E-06
Full X^2: 0.018444
Full R^2: 0.982059
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.
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.
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.
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.
ab51257 has been referenced in 45 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
- Hainfeld JF et al. Iodine nanoparticles enhance radiotherapy of intracerebral human glioma in mice and increase efficacy of chemotherapy. Sci Rep 9:4505 (2019). PubMed: 30872755
- Harmsen S et al. Detection of Premalignant Gastrointestinal Lesions Using Surface-Enhanced Resonance Raman Scattering-Nanoparticle Endoscopy. ACS Nano 13:1354-1364 (2019). PubMed: 30624916
- Kavita U et al. A Fit-for-Purpose Method for the Detection of Human Antibodies to Surface-Exposed Components of BMS-986263, a Lipid Nanoparticle-Based Drug Product Containing a siRNA Drug Substance. AAPS J 21:92 (2019). PubMed: 31332587