Human ICAM-1/CD54 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.	-Mouse (1:1000) --Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Upstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.
Protocol tips
-Mouse (1:1000) --Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Publication protocol

Cells were lysed in RIPA lysis buffer (1% NP-40, 50 mM Tris pH 7.4, 150 mM NaCl, 0.1% SDS, 1% sodium deoxycholate and 50 mM EDTA) with 1 × HALT protease and phosphatase inhibitors (Thermo Scientific). Lysates were run on SDS–polyacrylamide gel electrophoresis gels and transferred to nitrocellulose membranes. Membranes were immunoblotted with anti-ICAM-1 BBIG-I1 (1:1,000), VCAM-1 BBIG-V1 (1:1,000), E-selectin BBIG-E4 (1:1,000) (R&D systems), MAP4K4 A301-503A (1:2,000) (Bethyl), VE-cadherin 2500 (1:2,000), p-JNK1/2 4668 (1:1,000), total JNK1/2 9258 (1:1,000), p-p38 MAPK 4631 (1:1,000), p-IκBα 9246 (1:1,000), IκBα 4814 (1:1,000), p-p65 3039 (1:1,000), p65 8242 (1:1,000), HGK 3485 (1:1,000), p-Erk 4370 (1:1,000) (Cell Signaling Technology), total p38 MAPK sc-535 (1:1,000), total Erk1 sc-93 (1:4,000) (Santa Cruz) or lamin-β1 ab16048 (1:1,000) (Abcam) antibodies.

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Papers

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Manufacturer protocol

Download the product protocol from R&D Systems for Human ICAM-1/CD54 Antibody below.

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Videos

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