Confocal immunofluorescent analysis of HeLa cells using Calnexin (C5C9) Rabbit mAb (Alexa Fluor® 555 Conjugate) (red) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) #8844 (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Calnexin (C5C9) Rabbit mAb #2679.
|Immunofluorescence (Immunocytochemistry)||1:100 - 1:400|
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
NOTE: Cells should be grown, treated, fixed and stained directly in multiwell plates, chamber slides or on coverslips.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised December 2010
Protocol Id: 220
Calnexin (C5C9) Rabbit mAb (Alexa Fluor® 555 Conjugate) detects endogenous levels of total calnexin protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human calnexin.
Secretory and transmembrane proteins are synthesized on polysomes and translocate into the endoplasmic reticulum (ER) where they are often modified by the formation of disulfide bonds, amino-linked glycosylation and folding. To help proteins fold properly, the ER contains a pool of molecular chaperones including calnexin. Calnexin was first identified as being involved in the assembly of murine class I histocompatibility molecules (1,2). Calnexin is a calcium-binding protein embedded in the ER membrane that retains the newly synthesized glycoproteins inside the ER to ensure proper folding and quality control (3-5). The specificity of calnexin for a subset of glycoproteins is defined by a lectin site, which binds an early oligosaccharide intermediate on the folding glycoprotein (5).
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