Anti-CD58 antibody [EPR24012-2] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

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

• WB

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Western blot - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

This data was produced using ab308435, the same antibody clone but in a different formulation. 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] (<a href='/en-us/products/primary-antibodies/cd58-antibody-epr24012-2-ab308435'>ab308435</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) 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

false

• WB

Supplier Data

Western blot - Anti-CD58 antibody [EPR24012-2] - BSA and Azide free (AB308436)

This data was produced using ab308435, the same antibody clone but in a different formulation. 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] (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 ab265947 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L 680RD (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] (<a href='/en-us/products/primary-antibodies/cd58-antibody-epr24012-2-ab308435'>ab308435</a>) 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

Lane 4:

A-204 (human muscle rhabdomyosarcoma cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 28 kDa

Observed band size: 55-70 kDa

false