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Knockout Tested Rabbit Recombinant Monoclonal CD58 antibody. Suitable for WB, IHC-P and reacts with Human samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (AB308435), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (AB308435), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (AB308435), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (AB308435), expandable thumbnail
  • Western blot - Anti-CD58 antibody [EPR24012-2] (AB308435), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PICC/IFFlow CytIP
Human
Tested
Tested
Not recommended
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Target data

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.

Alternative names

LFA3, CD58, LFA3, Lymphocyte function-associated antigen 3, Ag3, Surface glycoprotein LFA-3

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal CD58 antibody. Suitable for WB, IHC-P and reacts with Human samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR24012-2

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD58 also known as lymphocyte function-associated antigen 3 (LFA-3) is a glycoprotein with a molecular mass of approximately 60 kDa. It is present on the surface of various cell types including antigen-presenting cells epithelial cells and some other leukocytes. This protein serves as a ligand for CD2 which is expressed on T cells and natural killer cells. Through its interaction with CD2 CD58 mediates cell adhesion and plays a role in immunological synapse formation which is essential for effective immune responses.

Biological function summary

Lymphocyte function-associated antigen 3 acts as a significant participant in the immune system. It plays a role in T cell activation and facilitates interactions between T cells and antigen-presenting cells. CD58 forms part of a complex network of cell surface proteins that enhances immune surveillance and response. Its role highlights its importance in adaptive immunity by promoting cell-to-cell communication and activation.

Pathways

CD58 participates in the cell adhesion pathways. It interacts closely with CD2 which is involved in the activation and signaling of T cells. Through this interaction CD58 plays roles in pathways like T cell receptor signaling and immune response. It enhances cell-cell adhesion helping T cells maintain their position during immune surveillance and activation.

Associated diseases and disorders

CD58 relates to conditions such as autoimmune diseases and certain cancers. In autoimmune disorders aberrant expression or function of CD58 may contribute to inappropriate immune activation leading to tissue damage. In cancer CD58 alterations can affect tumor immune evasion influencing how the immune system recognizes and attacks cancer cells. In multiple sclerosis an autoimmune disease the interaction between CD58 and CD2 has been studied providing insights into its role in disease mechanisms.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

7 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Immunohistochemical analysis of paraffin-embedded Human hepatocellular tissue labelling CD58 with ab308435 at 1/1000 (0.504 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human hepatocellular carcinoma.The section was incubated with ab308435 for 30 mins at room temperature. The
    immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Immunohistochemical analysis of paraffin-embedded Human esophagus tissue labelling CD58 with ab308435 at 1/1000 (0.504 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human esophagus (PMID: 1702102).The section was incubated with ab308435 for 30 mins at room temperature. The
    immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Immunohistochemical analysis of paraffin-embedded Human endometrioid c tissue labelling CD58 with ab308435 at 1/1000 (0.504 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human endometrioid carcinoma.The section was incubated with ab308435 for 30 mins at room temperature. The
    immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa ( tissue labelling CD58 with ab308435 at 1/500 (1.008 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) Wild-type HeLa cell pellet, no staining on (B) CD58 knockout HeLa cell pellet. The section was incubated with ab308435 for 30 mins at room temperature.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundThe immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 19268704; PMID: 3309127) and due to the high glycosylation of the protein.

    Exposure time: Lane 1: 92 seconds.; Lanes 2-5: 37 seconds.

    All lanes: Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435) at 1/1000 dilution

    Lane 1: Human kidney tissue lysate at 20 µg

    Lane 2: Human tonsil tissue lysate at 20 µg

    Lane 3: Human placenta tissue lysate at 20 µg

    Lane 4: Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg

    Lane 5: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

    Secondary

    Lanes 1 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2000 dilution

    Lanes 1 - 5: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

    Predicted band size: 28 kDa

    Observed band size: 55-70 kDa

  • Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lysates at 20 µg per lane.

    The samples were run on a Bis-Tris gel under reducing conditions.

    Western blot: Anti-CD58 antibody (ab308435) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20, 000 dilution, shown in red.

    In Western blot, ab308435 was shown to bind specifically to CD58. Target of interest was observed at 55-70 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in CD58 knockout cell line Human CD58 knockout HeLa cell line ab265947 (knockout cell lysate Human CD58 knockout HeLa cell lysate ab257880)(lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L 680RD (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

    Low expression: A-204 (PMID: 26039213).

    All lanes: Western blot - Anti-CD58 antibody [EPR24012-2] (ab308435) at 1/1000 dilution

    Lane 1: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: CD58 knockout HeLa whole cell lysate at 20 µg

    Lane 3: Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

    Secondary

    Lanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution

    Predicted band size: 28 kDa

    Observed band size: 55-70 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] (ab308435)

    Immunohistochemical analysis of paraffin-embedded Human follicular lym tissue labelling CD58 with ab308435 at 1/1000 (0.504 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human follicular lymphoma.The section was incubated with ab308435 for 30 mins at room temperature. The
    immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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