Anti-CD59 antibody [MEM-43/5] (ab9183)
Key features and details
- Mouse monoclonal [MEM-43/5] to CD59
- Suitable for: ICC/IF, IP, IHC-P, Flow Cyt, WB
- Reacts with: Mouse, Human
- Isotype: IgG2b
Overview
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Product name
Anti-CD59 antibody [MEM-43/5]
See all CD59 primary antibodies -
Description
Mouse monoclonal [MEM-43/5] to CD59 -
Host species
Mouse -
Specificity
CD59 antigen (human). MEM-43/5 reacts with well defined epitope (around L33) and does not compete with MEM-43 and many other CD59 antibodies -
Tested applications
Suitable for: ICC/IF, IP, IHC-P, Flow Cyt, WBmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Tissue, cells or virus corresponding to Human CD59. Thymocytes and T lymphocytes
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Epitope
The antibody MEM-43/5 reacts with well defined epitope around L33 (see Bodian et al) -
Positive control
- Flow cyt: blood Jeg3 cell line IF/ICC
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purity >95% by SDS-PAGE. -
Clonality
Monoclonal -
Clone number
MEM-43/5 -
Myeloma
unknown -
Isotype
IgG2b -
Light chain type
unknown -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab9183 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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ICC/IF |
Use a concentration of 5 µg/ml.
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IP |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use a concentration of 5 µg/ml.
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Flow Cyt |
Use a concentration of 0.5 - 4 µg/ml.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 14 kDa.
CD59 is GPI-anchored, so we recommend to use a laurylmatoside based lysis buffer or triton base buffer (see Bodian et al; 1% Triton X-100, 1 µg/ml leupeptin, 1 µg/ml pepstatin A and 1 mM phenlymethylsulphonyl fluoride in PBS), not NP40. |
Notes |
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ICC/IF
Use a concentration of 5 µg/ml. |
IP
Use at an assay dependent concentration. |
IHC-P
Use a concentration of 5 µg/ml. |
Flow Cyt
Use a concentration of 0.5 - 4 µg/ml. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 14 kDa. CD59 is GPI-anchored, so we recommend to use a laurylmatoside based lysis buffer or triton base buffer (see Bodian et al; 1% Triton X-100, 1 µg/ml leupeptin, 1 µg/ml pepstatin A and 1 mM phenlymethylsulphonyl fluoride in PBS), not NP40. |
Target
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Function
Potent inhibitor of the complement membrane attack complex (MAC) action. Acts by binding to the C8 and/or C9 complements of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. Involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase.
The soluble form from urine retains its specific complement binding activity, but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes. -
Involvement in disease
Defects in CD59 are the cause of CD59 deficiency (CD59D) [MIM:612300]. -
Sequence similarities
Contains 1 UPAR/Ly6 domain. -
Post-translational
modificationsN- and O-glycosylated. The N-glycosylation mainly consists of a family of biantennary complex-type structures with and without lactosamine extensions and outer arm fucose residues. Also significant amounts of triantennary complexes (22%). Variable sialylation also present in the Asn-43 oligosaccharide. The predominant O-glycans are mono-sialylated forms of the disaccharide, Gal-beta-1,3GalNAc, and their sites of attachment are probably on Thr-76 and Thr-77. The GPI-anchor of soluble urinary CD59 has no inositol-associated phospholipid, but is composed of seven different GPI-anchor variants of one or more monosaccharide units. Major variants contain sialic acid, mannose and glucosamine Sialic acid linked to an N-acetylhexosamine-galactose arm is present in two variants.
Glycated. Glycation is found in diabetic subjects, but only at minimal levels in nondiabetic subjects. Glycated CD59 lacks MAC-inhibitory function and confers to vascular complications of diabetes. -
Cellular localization
Cell membrane. Secreted. Soluble form found in a number of tissues. - Information by UniProt
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Database links
- Entrez Gene: 966 Human
- Entrez Gene: 12509 Mouse
- Omim: 107271 Human
- SwissProt: P13987 Human
- SwissProt: O55186 Mouse
- Unigene: 278573 Human
- Unigene: 709466 Human
- Unigene: 710641 Human
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Alternative names
- 16.3A5 antibody
- 1F5 antibody
- 1F5 antigen antibody
see all
Images
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Immunohistochemistry parafin embedded sections staining of huam skin tissue using ab9183.
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Flow cytometric analysis of Human Peripheral Blood cells labelling CD59 with ab9183 at 0.6 ug/ml showing separation of human neutrophil granulocytes (red-filled) from human CD59 negative blood debris (black-dashed).
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ICC/IF image of ab9183 stained Jeg3 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9183, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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IHC image of ab9183 staining CD59 in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab9183, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Overlay histogram showing Jurkat cells stained with ab9183 (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 (ab9183, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab9183 has been referenced in 13 publications.
- Okutomi T et al. HIV Reactivation in Latently Infected Cells With Virological Synapse-Like Cell Contact. Viruses 12:N/A (2020). PubMed: 32276457
- Davids M et al. Homozygous splice-variants in human ARV1 cause GPI-anchor synthesis deficiency. Mol Genet Metab 130:49-57 (2020). PubMed: 32165008
- Liu J et al. Single-cell RNA sequencing study of retinal immune regulators identified CD47 and CD59a expression in photoreceptors-Implications in subretinal immune regulation. J Neurosci Res 98:1498-1513 (2020). PubMed: 32166783
- Aitken RJ et al. Patterns of MTT reduction in mammalian spermatozoa. Reproduction 160:431-445 (2020). PubMed: 32567557
- Zhou Y et al. CD59 is a potential biomarker of esophageal squamous cell carcinoma radioresistance by affecting DNA repair. Cell Death Dis 9:887 (2018). PubMed: 30166523