Anti-CD58 antibody [TS2/9] (ab171087)
Key features and details
- Mouse monoclonal [TS2/9] to CD58
- Suitable for: ICC/IF, Flow Cyt, WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG1
Overview
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Product name
Anti-CD58 antibody [TS2/9]
See all CD58 primary antibodies -
Description
Mouse monoclonal [TS2/9] to CD58 -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, Flow Cyt, WBmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Full length protein corresponding to Human CD58. Native protein
Database link: P19256 -
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
Constituent: 99% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
TS2/9 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab171087 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 |
1/10 - 1/100.
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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WB |
1/100 - 1/500. Predicted molecular weight: 24 kDa.
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Notes |
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ICC/IF
1/10 - 1/100. |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
1/100 - 1/500. Predicted molecular weight: 24 kDa. |
Target
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Function
Ligand of the T-lymphocyte CD2 glycoprotein. This interaction is important in mediating thymocyte interactions with thymic epithelial cells, antigen-independent and -dependent interactions of T-lymphocytes with target cells and antigen-presenting cells and the T-lymphocyte rosetting with erythrocytes. In addition, the LFA-3/CD2 interaction may prime response by both the CD2+ and LFA-3+ cells. -
Sequence similarities
Contains 1 Ig-like C2-type (immunoglobulin-like) domain. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 965 Human
- Omim: 153420 Human
- SwissProt: P19256 Human
- Unigene: 34341 Human
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Alternative names
- AG3 antibody
- CD58 antibody
- CD58 antigen (lymphocyte function associated antigen 3)1 antibody
see all
Images
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Flow cytometry analysis of CD58 showing positive staining in the membrane of PBMC cells compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 ul of cell solution was added to each tube at a dilution of 2x10^7 cells/ml, followed by the addition of 50 ul of isotype control and ab171087 (0.5 ug/test). Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4ºC in the dark. FACS analysis was performed using 400 ul of cell buffer.
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All lanes : Anti-CD58 antibody [TS2/9] (ab171087) at 1/250 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CD58 knockout HAP1 whole cell lysate
Lane 3 : THP1 whole cell lysate
Lane 4 : Raji whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 24 kDaLanes 1 - 4: Merged signal (red and green). Green - ab171087 observed at 43 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab171087 was shown to specifically react with CD58 in wild-type HAP1 cells as signal was lost in CD58 knockout cells. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. Ab171087 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
Immunofluorescent analysis of CD58 (green) showing staining in the cytoplasm of Raji cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD58 monoclonal antibody (ab171087) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
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Flow cytometry analysis of CD58 showing positive staining in the membrane of PBMC cells. Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 ul of cell solution was added to each tube at a dilution of 2x10^7 cells/ml, followed by the addition of 50 ul of isotype control and ab171087 (0.5 ug/test). Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4ºC in the dark. FACS analysis was performed using 400 ul of cell buffer.
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Flow cytometry analysis of CD58 showing weakly positive staining in the membrane of BAF-3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with ab171087 (0.25 ug/test) for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
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All lanes : Anti-CD58 antibody [TS2/9] (ab171087) at 1/100 dilution
Lane 1 : Raji cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : BAF-3 cell lysate
Lysates/proteins at 25 µg per lane.
Predicted band size: 24 kDa
Observed band size: 24 kDa
Protocols
Datasheets and documents
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Datasheet download
References (0)
ab171087 has not yet been referenced specifically in any publications.