Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1802Y] to Prion protein PrP
- Suitable for: WB, Flow Cyt, IHC-P
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Prion protein PrP antibody [EP1802Y]
See all Prion protein PrP primary antibodies -
Description
Rabbit monoclonal [EP1802Y] to Prion protein PrP -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-P MouseRatHumanWB MouseRatHuman -
Immunogen
Synthetic peptide within Human Prion protein PrP aa 200-300 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: Human, rat and mouse brain tissue lysates. SK-MEL-28 and Neuro-2a whole cell lysates. ab74056; IHC-P: brain glioma tissue, Human, mouse and rat cerebrum tissue; Flow Cyt: SH-SY5Y cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1802Y -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Prion protein PrP antibody [EP1802Y] (ab52604) at 1/5000 dilution (Purified)
Lane 1 : Human brain lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat brain lysate
Lane 4 : SK-MEL-28 (Human malignant melanoma ) whole cell lysate
Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 28 kDa
Observed band size: 20-37 kDa why is the actual band size different from the predicted?The molecular weights observed represent different glycosation states and are consistent with what has been described in the literature (PMID: 20670940, PMID: 19568430, PMID: 15240877 and PMID: 22558368).
Blocking/Diluting buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells, labeling Prion protein PrP with Purified ab52604 at 1:70 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Anti-Prion protein PrP antibody [EP1802Y] (ab52604) at 1/5000 dilution (unpurified) +
Recombinant Mouse Prion protein PrP (ab74056) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 28 kDa
Exposure time: 10 secondsThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab52604 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Overlay histogram showing SH-SY5Y cells stained with unpurified ab52604 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52604, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of brain glioma using ab52604 (unpurified) at a dilution of 1/100. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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