Recombinant Anti-CD99 antibody [EPR3097Y] (ab75858)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3097Y] to CD99
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD99 antibody [EPR3097Y]
See all CD99 primary antibodies -
Description
Rabbit monoclonal [EPR3097Y] to CD99 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP-1 lysates. IHC-P: human prostatic hyperplasia, Ewing's sarcoma and tonsil tissues. ICC/IF: Jurkat cells. IP: MOLT-4 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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3097Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75858 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
1/50.
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WB |
1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 19 kDa).
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IP |
1/20.
For unpurified use at 1/100 |
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IHC-P |
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
1/50. |
WB
1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 19 kDa). |
IP
1/20. For unpurified use at 1/100 |
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Involved in T-cell adhesion processes. It is involved in spontaneous rosette formation with erythrocytes. -
Sequence similarities
Belongs to the CD99 family. -
Post-translational
modificationsExtensively O-glycosylated. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 4267 Human
- Omim: 313470 Human
- SwissProt: P14209 Human
- Unigene: 653349 Human
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Alternative names
- 12E7 antibody
- Antigen identified by monoclonal 12E7, Y homolog antibody
- Antigen identified by monoclonal antibodies 12E7, F21 and O13 antibody
see all
Images
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Anti-CD99 antibody [EPR3097Y] (ab75858) at 1/1000 dilution (Purified) + THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 19 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CD99 with purified ab75858 at 1/50 dilution (2.8 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
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ab75858 (Purified) at 1:20 dilution (0.6 µg) immunoprecipitating CD99 in MOLT-4 whole cell lysate.
Lane 1 (input): MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate 10 µg
Lane 2 (+): ab75858 & MOLT-4 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab75858 in MOLT-4 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling CD99 with Purified ab75858 at 1:1000 dilution (0.14 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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ab75858 at 1/1000 dilution staining CD99 in human Ewing's sarcoma by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of Jurkat (Human acute T cell leukemia) labeling CD99with purified ab75858at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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ab75858 at 1/1000 dilution staining CD99 in human tonsil by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab75858 has been referenced in 2 publications.
- Conn E et al. Development of an Ewing sarcoma cell line with resistance to EWS-FLI1 inhibitor YK-4-279. Mol Med Rep 21:1667-1675 (2020). PubMed: 32016454
- Chung SS et al. CD99 is a therapeutic target on disease stem cells in myeloid malignancies. Sci Transl Med 9:N/A (2017). WB ; Human . PubMed: 28123069