Anti-CD62E antibody [RM2070]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver. The section was incubated with ab322206 for 30 mins at room temperature. The 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.

• Flow Cyt

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Flow Cytometry - Anti-CD62E antibody [RM2070] (AB322206)

Flow cytometric analysis of HUVEC (human umbilical vein endothelial cell) treated with 20ng/ml Interleukin 1 beta for 4 hours (Right) / Untreated control (Left) cells labeling CD62E with ab322206 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessel of human colon carcinoma. The section was incubated with ab322206 for 30 mins at room temperature. The 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-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human CD62E expression vector containing a Myc-His tag. (B) HEK-293T transfected with a human CD62L expression vector containing a Myc-His tag. (C) HEK-293T transfected with a human CD62P expression vector containing a Myc-His tag. (D) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling CD62E with ab322206 at 1/2000 (0.244 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on HEK-293T transfected with a Myc-His-tagged human CD62E construct (image A). No staining on HEK-293T transfected with a Myc-His-tagged human CD62L construct (image B), human CD62P construct (image C). No staining on HEK-293T transfected with empty plasmid (image D). The section was incubated with ab322206 for 30 mins at room temperature. The 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD62E antibody [RM2070] (AB322206)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling CD62E with ab322206 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membrane and cytoplasmic staining in HUVEC cell line (shown in green) treated with IL-1beta (20 ng/mL) for 4 hr. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Human appendicitis tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessel of human appendicitis. The section was incubated with ab322206 for 30 mins at room temperature. The 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-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessel of human tonsil. The section was incubated with ab322206 for 30 mins at room temperature. The 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.

• IP

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Immunoprecipitation - Anti-CD62E antibody [RM2070] (AB322206)

All lanes:

Immunoprecipitation - Anti-CD62E antibody [RM2070] (ab322206) at 1/30 dilution

All lanes:

HUVEC (human umbilical vein endothelial cell) treated with 20ng/ml IL-lbeta for 4h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Mouse Crohn's disease model tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessel of mouse Crohn's disease model. The section was incubated with ab322206 for 30 mins at room temperature. The 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse cerebrum. The section was incubated with ab322206 for 30 mins at room temperature. The 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody [RM2070] (AB322206)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CD62E with ab322206 at 1/100 (4.88 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse liver. The section was incubated with ab322206 for 30 mins at room temperature. The 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD62E antibody [RM2070] (AB322206)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CT26.WT (mouse colon fibroblast) cells labelling CD62E with ab322206 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membrane and cytoplasmic staining in CT26.WT cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Negative control : Neuro-2a. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-CD62E antibody [RM2070] (AB322206)

All lanes:

Immunoprecipitation - Anti-CD62E antibody [RM2070] (ab322206) at 1/30 dilution

All lanes:

CT26.WT (mouse colorectal carcinoma fibroblast) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CD62E antibody [RM2070] (AB322206)

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

Exposure Time Lane 1 : 180s, Lane 2 : 40s

Compared with ab300557, ab319105 and ab322206 have higher sensitivity, we recommend ab319105 and ab322206 as an alternative for testing CD62E in western blot.

Lanes 1 - 2:

Anti-CD62E antibody [EPR25616-62] (<a href='/en-us/products/primary-antibodies/cd62e-antibody-epr25616-62-ab300557'>ab300557</a>) at 1/1000 dilution

Lane 3:

Western blot - Anti-CD62E antibody [EPR28605-81] (<a href='/en-us/products/primary-antibodies/cd62e-antibody-epr28605-81-ab319105'>ab319105</a>) at 1/1000 dilution

Lane 4:

Western blot - Anti-CD62E antibody [RM2070] (ab322206) at 1/1000 dilution

All lanes:

HUVEC (human umbilical vein endothelial cell) treated with 20ng/ml IL- lbeta for 4h whole cell lysate at 20 µg

Secondary

All lanes:

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 1/20000 dilution

Observed band size: 100 kDa,115 kDa

false

• WB

Supplier Data

Western blot - Anti-CD62E antibody [RM2070] (AB322206)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 7589110).

Negative control : Neuro-2a.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 15 seconds; Lanes 3-4 : 81 seconds.

All lanes:

Western blot - Anti-CD62E antibody [RM2070] (ab322206) at 1/1000 dilution

Lane 1:

Untreated HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

HUVEC treated with 20ng/ml IL-lbeta for 4h whole cell lysate at 20 µg

Lane 3:

CT26.WT (mouse colorectal carcinoma fibroblast) whole cell lysate at 20 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Secondary

All lanes:

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 1/20000 dilution

Observed band size: 100 kDa,115 kDa,36 kDa

false