Calnexin Monoclonal Antibody (AF18)

货号：MA3-027

规格：100 µL

价格：5439

产品类型：流式抗体

品牌：Thermo Fisher

物种：人/小鼠

宿主：小鼠

抗体亚型：其它

荧光染料：其它

类型:	单抗	同型对照：	IgG1
免疫原性：	Synthetic peptide corresponding to residues C T(643) G E E D T S E K D E L(654) of rat GRP94	用法：	1:50-1:500(IF);1:50-1:500/mL(ICC);1:20(IHC);5 µg(IP);1:100-1:1000(WB)

Product Specific InformationMA3-027 detects calnexin from human and mouse tissues.MA3-027 has been successfully used in Western blot, immunocytochemistry, immunofluorescence and immunoprecipitation protocols. By Western blot, this antibody detects an ~67 kDa band representing calnexin in human liver, skin, adrenal gland, heart, colon, testes and ovary extracts. This antibody is not recommended for human kidney or mouse liver lysates in Western blot applications. Immunocytochemical staining of calnexin in Huh-7 cells with MA3-027 yields a pattern consistent with specific endoplasmic reticulum staining.The MA3-027 antigen is Focus human hepatoma cell lysate.Target InformationCalnexin, also referred to as IP90, p88 and p90, is an ~90 kDa integral membrane protein of the endoplasmic reticulum (ER). Many resident ER proteins act as molecular chaperones and participate in the proper folding of polypeptides and their assembly into multisubunit proteins. Studies indicate that calnexin associates with the major histocompatibility complex (MHC) class I heavy chains, partial complexes of the T cell receptor and B cell membrane immunoglobulin, but not with completed receptor complexes. It has been shown that calnexin is a chaperone that retains incompletely or improperly folded proteins in the ER.The sequence Lys-Asp-Glu-Leu (KDEL) or a closely related sequence, is present at the carboxy-terminus of soluble ER resident proteins such as GRP78 and GRP94 and protein disulfide isomerase. Integral membrane ER resident proteins, like calnexin, often lack this KDEL sequence but contain positively charged cytosolic residues that ensure ER retention. Calnexin contains a large ER luminal domain (461 amino acids), a transmembrane segment (22 amino acids), and a cytoplasmic tail (89 amino acids).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

Calnexin Antibody (MA3-027) in IHCImmunohistochemistry was performed on normal deparaffinized Human kidney tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Anti-Calnexin (Product # MA3-027) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting. Calnexin Antibody (MA3-027) in IFImmunofluorescent analysis of calnexin (red) in U87 cells. Cell fixed with 4% paraformaldehyde were permeabilized with 0.05% Triton X-100 in PBS for 60 seconds at room temperature and blocked with 5% normal donkey serum for 30 minutes at room temperature. Cells were probed with a calnexin monoclonal antibody (Product # MA3-027) at a dilution of 1:100 for 2 hours at room temperature, washed with PBS, and incubated with a fluorescently-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:100 for 45 minutes at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a fluorescent microscope at 40X magnification. Data courtesy of the Innovators Program. Calnexin Antibody (MA3-027) in IPImmunoprecipitation of CANX was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 ug whole cell lysate with 5 ug of monoclonal CANX antibody (Product # MA3-027) rotating 60 min at RT. The immune complexes were captured on 625 ug of Dynabeads M-280 sheep anti-mouse IgG (Product # 11202D), washed extensively, and eluted in NuPAGE LDS sample buffer (Product # NP0007). Samples were resolved onto a 4-12% Bis-Tris protein gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to nitrocellulose membrane (Product # IB23001) and blocked in 5% milk. CANX was detected using a monoclonal CANX antibody (Product # MA3-027) at a dilution of 1:2000, followed by incubation with anti-mouse secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

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参考文献：

1.The Journal of neuroscience : the official journal of the Society for NeuroscienceDisturbed neuronal ER-Golgi sorting of unassembled glycine receptors suggests altered subcellular processing is a cause of human hyperekplexia."MA3-027 was used in Immunocytochemistry to conclude that a proportion of GlyR α1 mutants can be transported to the plasma membrane but do not necessarily form functional ion channels."AuthorsSchaefer N,Kluck CJ,Price KL,Meiselbach H,Vornberger N,Schwarzinger S,Hartmann S,Langlhofer G,Schulz S,Schlegel N,Brockmann K,Lynch B,Becker CM,Lummis SC,Villmann C2.OncotargetThe anti-tumor drug 2-hydroxyoleic acid (Minerval) stimulates signaling and retrograde transport."MA3027 was used in immunocytochemistry to show that 2-hydroxyoleic acid treatment alters cell signaling and intracellular transport"AuthorsTorgersen ML,Klokk TI,Kavaliauskiene S,Klose C,Simons K,Skotland T,Sandvig K3.EMBO reportsRAB2A controls MT1-MMP endocytic and E-cadherin polarized Golgi trafficking to promote invasive breast cancer programs."MA3-027 was used in immunocytochemistry to analyze control of MT1-MMP endocytic and E-cadherin polarized golgi trafficking to promote invasive breast cancer programs by RAB2A"AuthorsKajiho H,Kajiho Y,Frittoli E,Confalonieri S,Bertalot G,Viale G,Di Fiore PP,Oldani A,Garre M,Beznoussenko GV,Palamidessi A,Vecchi M,Chavrier P,Perez F,Scita G4.The Journal of neuroscience : the official journal of the Society for NeuroscienceDisturbed neuronal ER-Golgi sorting of unassembled glycine receptors suggests altered subcellular processing is a cause of human hyperekplexia."MA3-027 was used in Immunocytochemistry to conclude that a proportion of GlyR α1 mutants can be transported to the plasma membrane but do not necessarily form functional ion channels."AuthorsSchaefer N,Kluck CJ,Price KL,Meiselbach H,Vornberger N,Schwarzinger S,Hartmann S,Langlhofer G,Schulz S,Schlegel N,Brockmann K,Lynch B,Becker CM,Lummis SC,Villmann C5.The Journal of neuroscience : the official journal of the Society for NeuroscienceDisturbed neuronal ER-Golgi sorting of unassembled glycine receptors suggests altered subcellular processing is a cause of human hyperekplexia."MA3-027 was used in Immunocytochemistry to conclude that a proportion of GlyR α1 mutants can be transported to the plasma membrane but do not necessarily form functional ion channels."AuthorsSchaefer N,Kluck CJ,Price KL,Meiselbach H,Vornberger N,Schwarzinger S,Hartmann S,Langlhofer G,Schulz S,Schlegel N,Brockmann K,Lynch B,Becker CM,Lummis SC,Villmann C

技术参数

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