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Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)
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Western blot - Calnexin antibody (ab22595)
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Western blot - Calnexin - ER membrane marker antibody (ab22595)
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Product Name
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Calnexin - ER membrane marker antibody
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See all Calnexin - ER membrane marker antibodies (10)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Calnexin - ER membrane marker
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Calnexin.
(Peptide available as ab23379.)
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Reacts with
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Hu, Ms, Rat, Mmst
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Predicted to react
(species key)
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Dog
(Due to sequence homology)
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Tested applications
(see key)
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ICC/IF, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC/IF: Use at a concentration of 5 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 90 kDa). Can be blocked with Calnexin peptide (ab23379).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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ab22595 gave a positive result in the following Human Whole Cell Lysates:
Hela, MCF-7
Mouse Whole Cell Lystates:
NIH 3T3, MEF1
Mouse Tissue Lysates:
Brain, Liver, Heart, Kidney, Pancreas, Testis, Skeletal muscle, Spinal cord, Ovary
Rat Whole Cell Lysates:
PC12,
Rat Tissue Lysates:
Brain, Liver, Heart, Kidney
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Cellular localization
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Endoplasmic reticulum
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Research areas
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Neuroscience >> Sensory System >> Visual system
Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
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Relevance
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Calnexin, 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 histocompatability 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 GRP 78 and GRP 94 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). These features distinguish calnexin from soluble ER chaperones that cannot interact with the transmembrane and cytosolic domains of integral membrane proteins. The amino acid sequence of calnexin is highly conserved among species, and shares regions of high sequence homology with calreticulin.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.10 - 0.30 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Store in the dark. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Calnexin - ER membrane marker antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Calnexin - ER membrane marker antibody images, references, products related to ab22595 and other tools.
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Calnexin - ER membrane marker antibody images:
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 Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)
ICC/IF image of ab22595 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22595, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Western blot - Calnexin antibody (ab22595)
All lanes : Calnexin - ER membrane marker antibody (ab22595) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line)
Lane 2 : U2OS Whole Cell Lysate
Lane 3 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) with Calnexin peptide (ab23379) at 1 µg/ml
Lane 5 : U2OS Whole Cell Lysate with Calnexin peptide (ab23379) at 1 µg/ml
Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Calnexin peptide (ab23379) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 90 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
Recent batches of ab22595 (AP217379 and AP151845) detect a band of ~ 75 kDa in Hela, U2OS and MCF-7 lysates. This band is completely blocked by the immunizing peptide so we believe this represents Calnexin. Moreoever, a band of the same size is detected by other Calnexin antibodies tested.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Calnexin - ER membrane marker antibody (ab22595)
Kidney cortex using ab22595 shows clear cytoplasmic staining patterns.
The visceral cells of the Glomerular tuft ( podocytes ) are strongly stained (indicated by red arrowheads). Distal convoluted tubular cells are generally moderately positive (with exceptions that are strongly positive). However, most of the cells that line the Proximal Convoluted Tubules (indicated by green arrowheads) are strongly positive. Carl Hobbs, King`s College London, United Kingdom
See Abreview
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References for Calnexin - ER membrane marker antibody (ab22595)
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This product has been used in: (two most recent references)
Wachten S et al. Distinct pools of cAMP centre on different isoforms of adenylyl cyclase in pituitary-derived GH3B6 cells. J Cell Sci 123:95-106 (2010). WB; Human. PubMed: 20016070
Takahashi Y et al. Identification of a novel palmitylation site essential for membrane association and isomerohydrolase activity of RPE65. J Biol Chem 284:3211-8 (2009). ICC/IF; Human. PubMed: 19049981
If you publish research using ab22595 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Calnexin - ER membrane marker
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Calnexin - ER membrane marker antibody - more information
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