Recombinant Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] (ab284040)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25045-51] to SARS-CoV-2 orf3d
- Suitable for: WB, Indirect ELISA, Flow Cyt (Intra), ICC/IF
Related conjugates and formulations
Overview
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Product name
Anti-SARS-CoV-2 orf3d antibody [EPR25045-51]
See all SARS-CoV-2 orf3d primary antibodies -
Description
Rabbit monoclonal [EPR25045-51] to SARS-CoV-2 orf3d -
Host species
Rabbit -
Tested applications
Suitable for: WB, Indirect ELISA, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: SARS-CoV-2 -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SARS-CoV2 Orf3d peptide (aa12-32) I-ELISA: SARS-CoV2 ORF3d peptide (aa 12-32) ICC/IF: HEK-293T cells transfected with a SARS-CoV2 orf3d expression vector containing a myc tag. Flow Cyt (Intra): HEK-293T cells transfected with a SARS-CoV2 orf3d expression vector containing GFP tag.
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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.
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
EPR25045-51 -
Isotype
IgG
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab284040 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 |
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WB |
1/1000.
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Indirect ELISA |
Use a concentration of 2e-005 - 1 µg/ml.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/100.
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Notes |
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WB
1/1000. |
Indirect ELISA
Use a concentration of 2e-005 - 1 µg/ml. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/100. |
Images
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All lanes : Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] (ab284040) at 1/1000 dilution
Lane 1 : Orf3d peptide (aa12-32) at 0.5 µg
Lane 2 : Orf3d peptide (aa12-32) at 0.2 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/20000 dilution
Observed band size: 6 kDa why is the actual band size different from the predicted?ab284040 was shown to bind specifically to SARS-CoV2 Orf3d peptide in Western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.
Blocking buffer: 5% BSA in TBS-0.1% Tween® 20 (TBS-T)
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labelling SARS-CoV-2 orf3d with primary antibody anti-SARS-CoV-2 orf3d (ab284040) at 1/100 dilution, followed by Goat Anti-Rabbit (Alexa Fluor® 594) (ab150089) (green) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in HEK-293T cells transfected with a SARS-CoV2 orf3d expression vector containing a myc tag, no staining in HEK-293T cells transfected with a SARS-CoV2 orf8 expression vector containing a myc tag. Myc-Tag Mouse mab (Alexa Fluor® 647) (red) was used to counterstain cells at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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Intracellular Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a SARS-CoV-2 orf8 (Left) expression vector containing a myc tag or SARS-CoV2 orf3d (Right) expression vector containing GFP tag labeling SARS-CoV-2 orf3d with ab284040 at 1/500 dilution (Left and Right). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 647, ab150083) secondary antibody was used at 1/5000 dilution.
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Indirect ELISA showing primary antibody ab284040 binding to SARS-CoV2 ORF3d peptide (aa 12-32) . Plates were coated with SARS-CoV2 ORF3d peptide (aa 12-32) and recombinant SARS-CoV2 ORF8 (aa 16-121) protein (His tag) at 1000 ng/ml. Binding of ab284040 was assessed in a serial dilution range 0.016- 1000 ng/mL (a 3-fold serial dilution).
Binding was detected using pre-adsorbed secondary antibody, goat anti-rabbit IgG H&L (HRP, ab97080) at 1/2000 dilution.
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
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SDS download
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Datasheet download
References (0)
ab284040 has not yet been referenced specifically in any publications.