Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Biotin Rabbit monoclonal [PEG-B-47b] to Polyethylene glycol
- Suitable for: IHC-P, Sandwich ELISA, WB, ELISA
- Conjugation: Biotin
Product nameBiotin Anti-Polyethylene glycol antibody [PEG-B-47b]
See all Polyethylene glycol primary antibodies
DescriptionBiotin Rabbit monoclonal [PEG-B-47b] to Polyethylene glycol
Tested applicationsSuitable for: IHC-P, Sandwich ELISA, WB, ELISAmore details
Chemical/ Small Molecule corresponding to Polyethylene glycol conjugated to keyhole limpet haemocyanin. KLH-PEG with a terminal methoxy group.
- WB: BSA-PEG5K. IHC-P: Human cerebral cortex tissue
The smallest PEG molecule which our Polyethylene glycol antibody (ab53449) can detect is PEG750. Based on customer feedback, the largest PEG molecule detected is PEG 40K. Additionally, ab53449 detects both linear and branched PEG based on our in-house ELISA testing.
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.
Learn more about PEG RabMAb® primary antibodies and kit - PEG RabMAb Product Portal Page
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.
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.
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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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, PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab53449 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified use at 1/50 - 1/100. Images of positive IHC with this monoclonal antibody, clone PEG-B-47b, are on the ab51257 datasheet.
|Sandwich ELISA||Use at an assay dependent concentration.|
|WB||1/50000. Predicted molecular weight: 2 kDa.
For unpurified use at 1/5000 - 1/20000.
|ELISA||Use a concentration of 1 µg/ml.
Coating plates for ELISA: 0 to 500 ng/ml.
For unpurified use at 0.5µg/ml, coating plates for ELISA: 5 to 10 ug/ml.
- PEG antibody
Direct ELISA antigen dose-response curve using purified ab53449. Antigen concentration of 0-500 ng/mL. A peroxidase-conjugated steptavidin (1/1000) was used.
Biotin conjugate (ab53449) available for this clone.
Sandwich ELISA antigen dose-response curve using purified ab53449.
Capture antibody - Mouse anti-IFN alpha 2b (4000 ng/ml).
Antigens - PEG-IFN alpha 2b (0-1000 ng/ml).
Detection - ab53449 (500 ng) anti-Polyethylene glycol antibody [PEG-B-47b] (Biotin) followed by peroxidase-conjugated streptravidin (1/1000).
Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449) at 1/50000 dilution (purified) + BSA-PEG5K at 0.02 µg
Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 2 kDa
Observed band size: 280 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue showing negative staining with purified ab53449 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
ELISA assay using 5 µg/mL Anti-AFP RabMAb as the capture antibody and 0.5 µg/mL Anti-PEG-47-Biotin RabMAb (unpurified ab53449) as the detection antibody. HRP-avidin (1/3000) used for anti-PEG-47 detection. This ELISA assay could detect YCA1017 peptide-PEG antigen at a concentration as low as 0.9 ng/mL.
Direct ELISA assay using 63 ng/mL of antigen (GCSF-PEG) to coat the plate. HRP-avidin (1/3000) was used for the detection of varying amounts of anti-PEG-47-Biotin (unpurified ab53449).
Lane 1 : Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449) at 1/5000 dilution (unpurified)
Lane 2 : Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449) at 1/10000 dilution (unpurified)
Lane 3 : Biotin Anti-Polyethylene glycol antibody [PEG-B-47b] (ab53449) at 1/20000 dilution (unpurified)
All lanes : BSA-PEG
Predicted band size: 2 kDa
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.
ab53449 has been referenced in 12 publications.
- Stock AA et al. Conformal Coating of Stem Cell-Derived Islets for ß Cell Replacement in Type 1 Diabetes. Stem Cell Reports 14:91-104 (2020). PubMed: 31839542
- Che L et al. Computer-assisted engineering of programmed drug releasing multilayer nanomedicine via indomethacin-mediated ternary complex for therapy against a multidrug resistant tumor. Acta Biomater N/A:N/A (2019). PubMed: 31344512
- Poirier N et al. FR104, an antagonist anti-CD28 monovalent fab' antibody, prevents alloimmunization and allows calcineurin inhibitor minimization in nonhuman primate renal allograft. Am J Transplant 15:88-100 (2015). PubMed: 25488654
- der Valk FM et al. Prednisolone-containing liposomes accumulate in human atherosclerotic macrophages upon intravenous administration. Nanomedicine N/A:N/A (2015). PubMed: 25791806
- Jen J et al. Pegvisomant bioavailability of single 30 mg/mL subcutaneous injection compared to two 15 mg/mL subcutaneous injections: a pharmacokinetic, safety and tolerability study. Growth Horm IGF Res 23:114-9 (2013). ELISA . PubMed: 23651793