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
Product nameAnti-CD58 antibody [TS2/9]
See all CD58 primary antibodies
DescriptionMouse monoclonal [TS2/9] to CD58
Tested applicationsSuitable for: ICC/IF, Flow Cyt, WBmore details
Species reactivityReacts with: Mouse, Human
Full length protein corresponding to Human CD58.
Database link: P19256
Previously labelled as LFA3.
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Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped 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 bufferConstituent: 99% PBS
Concentration information loading...
PurityProtein A purified
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.
|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.|
FunctionLigand 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 similaritiesContains 1 Ig-like C2-type (immunoglobulin-like) domain.
Cellular localizationCell membrane.
- Information by UniProt
- AG3 antibody
- CD58 antibody
- CD58 antigen (lymphocyte function associated antigen 3)1 antibody
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.
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 kDa
Lanes 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.
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.
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.
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
ab171087 has not yet been referenced specifically in any publications.