Anti calnexin

Manufactured by LifeSpan BioSciences

Anti-calnexin is an antibody that recognizes the protein calnexin, which is an integral membrane protein found in the endoplasmic reticulum of eukaryotic cells. Calnexin functions as a molecular chaperone, assisting in the folding and quality control of newly synthesized glycoproteins. The anti-calnexin antibody can be used to detect and study the expression and localization of calnexin in various biological samples.

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2 protocols using anti calnexin

Immunoblotting Assay for NLRP3 Inflammasome

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Anti-ASC (Cell Signaling Technology, 67824, 1:1,000), anti-NLRP3 (Cell Signaling Technology, 15101, 1:1,000), anti-cleaved IL-1β (Cell Signaling Technology, 52718, 1:1,000), anti-pro–IL-1β (Cell Signaling Technology, 31202, 1:1,000), anti-DGAT1 (Gene-Protein Link, 1:1,000), anti–β-actin (Cell Signaling Technology, 3700, 1:2,000), anti-pro–caspase-1/cleaved caspase-1 p20 (AdipoGen Life Sciences, AG-20B-0042, 1:1,000), anti-NEK7 (Santa Cruz, sc-50756, 1:1,000), anti-GSTO1 (Proteintech, 15124-1-AP, 1:1,000), anti-GSH (ViroGen, 101-A, 1:1,000), anti-calnexin (LifeSpan Biosciences, LS-B9772-200, 1:1,000), anti-FACL4 (ABclonal, A20414, 1:1,000), peroxidase-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch, 111-035-144, 1:5,000), anti-mouse IgG (Jackson ImmunoResearch, 111-035-146, 1:5,000), peroxidase-conjugated donkey anti-goat IgG (Abcam, ab97110, 1:5,000), and Alexa Fluor 488–conjugated chicken anti-goat IgG (Invitrogen, 21467, 1:1,000) were used in our experiments.

Li S., Wang L., Xu Z., Huang Y., Xue R., Yue T., Xu L., Gong F., Bai S., Wu Q., Liu J., Lin B., Zhang H., Xue Y., Xu P., Hou J., Yang X., Jin T., Zhou R., Lou J., Xu T, & Bai L. (2021). ASC deglutathionylation is a checkpoint for NLRP3 inflammasome activation. The Journal of Experimental Medicine, 218(9), e20202637.

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Western Blot Analysis of Immune Proteins

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Guanidine whole-cell lysates were precipitated using a ProteoExtract protein precipitation kit (Calbiochem). Proteins were re-dissolved in 2% SDS/Tris 200 mM pH8.5 with sonication. Protein concentration was measured by BCA (Pierce) then lysates were reduced with 100 mM DTT in loading buffer for 20 min at 65°C. Equal protein amounts were separated by SDS/PAGE using 4%–12% Bis-Tris polyacrylamide gels (Life Technologies), then transferred to PVDF membranes using the iBlot transfer system (Life Technologies). The following primary antibodies were used: anti-IFIT1 (clone 3G8, Lifespan), anti-IFIT3 (cat no. GTX112442, GeneTex), anti-MX1 (clone 2G12, GeneTex), anti-Calnexin (cat no. C142768, Lifespan), anti-MHC class I (clone HC-10, Abcam), anti-CD99 (cat no. ab108297, Abcam), anti-CLEC1A (cat no. AF1704, R and D systems), anti-FAT1 (cat no. HPA023882, Sigma), anti-PCDHGC3 (cat no. sc-134416, Santa Cruz), anti-IRF3 (clone FL-425, Santa Cruz), anti-NF-kB p50/105 (cat no. sc-7178, Santa Cruz), anti-p65/RELA (cat no. sc-109, Santa Cruz), anti-Pan-Akt (clone C67E7, Cell Signaling), anti-actin (cat no. A2066, Sigma). HRP-conjugated secondary antibodies were anti-rabbit (Cell Signaling), anti-mouse (Santa Cruz) or anti-goat (Santa Cruz). Reactive bands were detected by SuperSignal West Pico substrate (Thermo).

Weekes M.P., Tomasec P., Huttlin E.L., Fielding C.A., Nusinow D., Stanton R.J., Wang E.C., Aicheler R., Murrell I., Wilkinson G.W., Lehner P.J, & Gygi S.P. (2014). Quantitative Temporal Viromics: An Approach to Investigate Host-Pathogen Interaction. Cell, 157(6), 1460-1472.

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