Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, BBA16, ELISA - Cell Based, ELISA Development, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # BBIG-E4
View all E-Selectin/CD62E Primary Antibodies »

Product Specifications

Immunogen

Activated HUVEC human umbilical vein endothelial cells

Specificity

Detects E‑Selectin/CD62E in ELISAs and Western blots. In immunofluorescence experiments, does not stain COS cells transfected with cDNA for human ICAM-1, PECAM‑1, L-Selectin, P-Selectin, or VCAM-1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human E‑Selectin/CD62E Antibody

Detection of E-Selectin/CD62E in HUVECs by Flow Cytometry.

Human umbilical cord endothelial cells (HUVECs) were cultured for 6 hours in the presence of 25 ng/mL of rhTNF-alpha (210-TA, filled histogram) or rested (open histogram) and stained with Mouse anti-human E-selectin/CD62E (BBA16) followed by Goat anti-Mouse APC-conjugated secondary antibody (F0101B). Gates were set based on isotype control (MAB002, data not shown). Staining was performed using our Staining Membrane-associated Proteins protocol.

E-Selectin/CD62E antibody in HUVEC Human Cells by Immunocytochemistry (ICC).

E-Selectin/CD62E in HUVEC Human Cells.

E-Selectin/CD62E was detected in immersion fixed HUVEC human umbilical vein endothelial cells activated with TNF-a (Catalog # 210-TA-010) using Mouse Anti-Human E-Selectin/ CD62E Monoclonal Antibody (Catalog # BBA16) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

E‑Selectin/CD62E antibody in Human Brain by Immunohistochemistry (IHC-P).

E‑Selectin/CD62E in Human Brain.

E-Selectin/CD62E was detected in immersion fixed paraffin-embedded sections of human brain (occipital cortex) using 10 µg/mL Mouse Anti-Human E-Selectin/CD62E Monoclonal Antibody (Catalog # BBA16) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of E-Selectin/CD62E by Western Blot

Detection of E-Selectin/CD62E by Western Blot

Induction of MIR181B expression by methotrexate (MTX) or adenosine is adenosine receptor A3 (ADORA3) dependent.(A) Knockdown for adenosine receptors A1, A2A, A2B, and A3 in HUVECs was performed to analyze MIR181B expression. three biological replicates. One-way ANOVA. (B) MIR181B expression in HUVECs transfected with Ctl-siRNA or ADORA3 siRNA after treatment with MTX (10 µM) or Ad (50 µM) or (C) treatment with TNF-alpha (10 ng/ml) alone or in combination MTX (10 µM) or Ad (50 µM). Three biological replicates. One-way ANOVA and Unpaired two-tailed Student t test. (D) Western blot analyses of VCAM-1, ICAM-1, and E-Selectin expression in HUVECs transfected with Ctl-siRNA or ADORA3 siRNA in the presence of TNF-alpha (10 ng/ml) in combination with either MTX (10 µM) or Ad (50 µM). Three biological replicates. Unpaired two-tailed Student t test. (E) in the presence of miRNA negative control (NS-m) or MIR181B mimics (181b-m) stimulated with TNF-alpha (10 ng/ml) or in combination with MTX (10 µM) or Ad (50 µM). Please see Figure 2—source data 1. Three biological replicates. Unpaired two-tailed Student t test. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. n.s. indicated non significance. All values represent mean ± SEM.Figure 2—source data 1.Knockdown efficiency for adenosine receptor siRNAs.mRNA, and protein expression analysis for (A, B) ADORA3 siRNA, (C, D) ADORA1 siRNA, (E, F) ADORA2A siRNA and (G, H) ADORA2B siRNA compared to control siRNA in HUVECs transfected for 36 hr. (A–H), n = 3. ***p<0.001. All values represent mean ± SEM. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33416495), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of E-Selectin/CD62E by Western Blot

Detection of E-Selectin/CD62E by Western Blot

Methotrexate (MTX) and Ad repress TNF-alpha -induced pro-inflammatory genes without affecting primary MIR181A-1 and MIR181B-1 expression.(A) Western blot analysis of VCAM-1, ICAM-1, and E-Selectin in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after stimulation of TNF-alpha (10 ng/ml) for 8 hr. Quantification of n = 3 independent experiments. (B) Real-time qPCR analysis of VCAM-1, ICAM-1, and E-Selectin in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after treatment with TNF-alpha (10 ng/ml) for 4 hr. Real-time qPCR analysis of (C) primary transcript of MIR181A1 or (D) primary transcript of MIR181B-1 in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after treatment with TNF-alpha (10 ng/ml) for 4 hr. (A–D), n = 3–6. *p<0.05; **p<0.01; ***p<0.001; ***p<0.0001. All values represent mean ± SEM. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33416495), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human E‑Selectin/CD62E Antibody

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: Human umbilical cord endothelial cells (HUVECs) cultured for 6 hours in the presence of 25 ng/mL of rhTNF-alpha (Catalog # 210-TA)

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HUVEC human umbilical vein endothelial cells stimulated with TNF-alpha (Catalog # 210-TA-010)

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (occipital cortex) subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Immunoprecipitation

Pigott, R. et al. (1991) J. Immunol. 147:130.

Western Blot

1 µg/mL
Sample: Recombinant Human E-Selectin/CD62E Fc Chimera (Catalog # 724-ES) under non-reducing conditions only

Human E-Selectin/CD62E Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-10 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human E‑Selectin/CD62E Biotinylated Antibody (Catalog # BBA8)
  • Standard: Recombinant Human E-Selectin/CD62E Protein, CF (Catalog # ADP1)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 4 reviews rated 4.5 using BBA16 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: E-Selectin/CD62E

E-Selectin, also known as Endothelial Leukocyte Adhesion Molecule 1 (ELAM-1), and Leukocyte-Endothelial Cell Adhesion Molecule 2 (LECAM-2), has been designated CD62E. It is a cell-surface glycoprotein expressed by activated endothelium and it mediates the adhesion of blood neutrophils.

Alternate Names

CD62E, ELAM1, LECAM2, SELE

Entrez Gene IDs

6401 (Human); 20339 (Mouse); 25544 (Rat); 102143564 (Cynomolgus Monkey)

Gene Symbol

SELE

Additional E-Selectin/CD62E Products

Product Documents for Human E‑Selectin/CD62E Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human E‑Selectin/CD62E Antibody

For research use only

Citations for Human E‑Selectin/CD62E Antibody

Customer Reviews for Human E‑Selectin/CD62E Antibody (4)

4.5 out of 5
4 Customer Ratings
5 Stars
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4 Stars
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3 Stars
25%
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1 Stars
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Customer Images

Showing  1 - 4 的 4 reviews Showing All
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  • Human E-Selectin/CD62E Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Colon adenocarcinoma tissue
    Species: Human
    Verified Customer | Posted 11/28/2021
    Colon adenocarcinoma
    Human E‑Selectin/CD62E Antibody BBA16
  • Human E-Selectin/CD62E Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 08/18/2021
    Human E‑Selectin/CD62E Antibody BBA16
  • Human E-Selectin/CD62E Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Thyroid carcinoma
    Species: Human
    Verified Customer | Posted 08/11/2021
    Human E‑Selectin/CD62E Antibody BBA16
  • Human E-Selectin/CD62E Antibody
    Name: Michelle Chen
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 04/26/2017
    HUVECs were stimulated with LPS for activation

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