Overview

  • 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, Blocking, ELISA, IHC-Fr, Neutralising, Flow Cyt, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length protein corresponding to Human CD58.
    Database link: P19256

  • General notes

    Previously labelled as LFA3.

Properties

  • 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...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    TS2/9
  • Isotype
    IgG1
  • Research areas

Applications

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
ICC/IF 1/10 - 1/100.
Blocking Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
WB 1/100 - 1/500. Predicted molecular weight: 24 kDa.

Target

  • 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
  • Database links
  • Alternative names
    • AG3 antibody
    • CD58 antibody
    • CD58 antigen (lymphocyte function associated antigen 3)1 antibody
    • CD58 antigen antibody
    • CD58 antigen, (lymphocyte function associated antigen 3) antibody
    • CD58 molecule antibody
    • FLJ23181 antibody
    • FLJ43722 antibody
    • LFA 3 antibody
    • LFA-3 antibody
    • LFA3 antibody
    • LFA3_HUMAN antibody
    • Lymphocyte Function Associated Antigen Type 3 antibody
    • Lymphocyte function-associated antigen 3 antibody
    • Surface glycoprotein LFA 3 antibody
    • Surface glycoprotein LFA-3 antibody
    see all

Images

  • 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 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.

  • 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

References

ab171087 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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