PA5-47320

antibody from Invitrogen Antibodies

Targeting: ICAM2 CD102

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Blocking/Neutralizing (Recommended by provider)

Antibody Data

Product number

PA5-47320

Provider

Invitrogen Antibodies

Product name

ICAM-2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) ICAM-1 and rhICAM-3 is observed. Reconstitute at 0.2 mg/mL in sterile PBS. Endoxin level is

Reactivity

Human

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.2 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ICAM-2 Polyclonal Antibody (Product # PA5-47320) and bands at 55-60 kDa corresponding to ICAM-2 was observed across all the cell lines tested except HeLa and HEK-293 which are reported to be negative. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), HUVEC (Lane 3), HeLa (Lane 4) and HEK-293 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.1 ug/ml) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014), 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of ICAM-2 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ICAM-2 Goat Polyclonal Antibody (Product # PA5-47320) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A27012) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.