Product nameAnti-Polyethylene glycol antibody [PEG-2-128]
See all Polyethylene glycol primary antibodies
DescriptionRabbit monoclonal [PEG-2-128] to Polyethylene glycol
Tested applicationsSuitable for: Sandwich ELISA, WB, ELISAmore details
Species reactivityReacts with: Species independent
Chemical/ Small Molecule corresponding to Polyethylene glycol conjugated to keyhole limpet haemocyanin. Synthetic peptide corresponds to Polyethylene glycol 20K (PEG20K).
EpitopeThis antibody recognizes the backbone of the Polyethylene glycol molecule. It can therefore be used to detect free forms of PEG. To detect conjugated forms of PEG, please see ab51257, which detects the terminal methoxy group of the PEG molecule.
Antibody Isotype: Please note, this RabMAb product's isotype is Rabbit IgM.
If you have any questions on our PEG products - please visit our Polyethylene glycol (PEG) FAQs page.
As this product's isotype is Rabbit IgM, KD values were provided by loading the antigen first before testing the antibody.
When compared to loading the antibody first and then the antigen, this method can give approximately 10-1 difference as determined by using ab51257 - Anti-Polyethylene glycol antibody [PEG-B-47] (of rabbit IgG isotype) as a control:
Antigen loaded first: 1.51E-10. Antibody loaded first: 2.41E-09.
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.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Dissociation constant (KD)KD = 8.24 x 10 -12 M Learn more about KD
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS, Tissue culture supernatant
Concentration information loading...
Purification notesHarvested from hybridoma is preliminarily purified via H2O dialysis and then centrifuged to remove some impurities components.
Primary antibody notesab133471 is useful in measuring the pharmacokinetics of Polyethylene glycol modified molecules.
Our Abpromise guarantee covers the use of ab133471 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
|WB||1/1000. The MW of the WB band for ab133471 varies based on the PEGylated molecule detected.|
|ELISA||Use a concentration of 2 - 10 µg/ml.|
- PEG antibody
Direct ELISA antigen dose-response curve using purified ab133471. Antigen concentration of 0-10000 ng/mL. A HRP-conjugated goat anti-rabbit IgM (1/1000) was used as the secondary antibody.
Direct ELISA antigen dose-response curve using purified ab133471. Antigen concentration of 0-100 ng/mL. A HRP-conjugated goat anti-rabbit IgM (1/1000) was used as the secondary antibody.
Sandwich ELISA antigen dose-response curve using purified ab133471.
Capture antibody - ab133471 (10000 ng/ml).
Antigens - MBP-PEG20K and ADI-PEG (0-1000 ng/ml).
Detection - ab53449 (500 ng) anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) followed by peroxidase-conjugated streptravidin (1/1000).
All lanes : Anti-Polyethylene glycol antibody [PEG-2-128] (ab133471) at 1/1000 dilution (purified)
Lane 1 : PEG20K
Lane 2 : MBP-PEG20K
Lysates/proteins at 5 µg per lane.
All lanes : Peroxidase-conjugated goat anti-rabbit IgM, (H+L) at 1/500 dilution
Blocking buffer: 15% BSA/TBS.
Dilution buffer: 10% PAA /TBST.
All lanes : Anti-Polyethylene glycol antibody [PEG-2-128] (ab133471) at 1/1000 dilution (unpurified)
Lane 1 : MBP-PEG20K at 1 µg
Lane 2 : MBP-PEG20K at 0.5 µg
Lane 3 : PEG20K at 1 µg
Lane 4 : PEG20K at 0.5 µg
All lanes : HRP-conjugated goat anti-rabbit IgM at 1/2000 dilution
Gel concentration: 4-20%
Various concentrations of Polyethylene glycol polymers with amine end groups were coated in 96 wells. 2 ug/ml of unpurified ab133471 was added. After washing, samples were incubated with 1 ug/ml of HRP-conjugated anti-rabbit IgM.
Various concentrations of Polyethylene glycol 20K (PEG20K) conjugated MBP and branched Polyethylene glycol conjugated BSA were coated in 96 wells. 2 ug/ml of unpurified ab133471 was added. After washing, samples were incubated with 1 ug/ml of HRP-conjugated anti-rabbit IgM.
10 ug/ml of unpurified ab133471 was coated into 96 wells. Serial dilution of Polyethylene glycol 20K (PEG20K) conjugated MBP and PEGylated ADI were added. After washing, 1 ug/ml of biotinylated anti-Polyethylene glycol antibody was added. Streptavidin-HRP was then used to develop the color.
Method：APS sensor + Antigen (PEG 5K-BSA at 0.3µg/ml)+ antibody (ab133471 at 0.0312, 0.0625, 0.125, 0.25, 0.5, 1 µg/m).
Sample ID: PEG-2-128
KD (M): 8.24E-12
kon Error: 3.59E+04
kdis Error: 6.46E-07
Full X^2: 6.0601
Full R^2: 0.9985
This product has been referenced in:
- Sanchez MD et al. Development and evaluation of a host-targeted antiviral that abrogates herpes simplex virus replication through modulation of arginine-associated metabolic pathways. Antiviral Res 132:13-25 (2016). WB . Read more (PubMed: 27192555) »
- Shiraishi K et al. Exploring the relationship between anti-PEG IgM behaviors and PEGylated nanoparticles and its significance for accelerated blood clearance. J Control Release 234:59-67 (2016). Sandwich ELISA . Read more (PubMed: 27164541) »