Recombinant 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, IHC-P
- Reacts with: Species independent
Related conjugates and formulations
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, WB, IHC-Pmore details -
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
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
-
Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
PEG-B-47 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
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.
|
|
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. |
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
Images
-
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 secondsBlocking/Diluting buffer and concentration 5% NFDM/TBST
-
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).
-
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 dilutionBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
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
-
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/mLSecondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.
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.
Datasheets and documents
-
SDS download
-
Datasheet download
References (51)
ab51257 has been referenced in 51 publications.
- Smith JP et al. Cholecystokinin-B Receptor-Targeted Nanoparticle for Imaging and Detection of Precancerous Lesions in the Pancreas. Biomolecules 11:N/A (2021). PubMed: 34944412
- 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