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|Antigen||Calnexin (CANX) Antibodies|
|Epitope||C-Term, AA 550-591 Alternatives|
|Reactivity||Hamster, Human, Mouse (Murine), Rat (Rattus) Alternatives|
|Conjugate||This Calnexin antibody is un-conjugated Alternatives|
Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Simple Western (SimWes), Western Blotting (WB)
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Product Details anti-Calnexin AntibodyTarget Details Calnexin Application Details Handling References for anti-Calnexin antibody (ABIN151474) Images
|Purification||Immunogen affinity purified|
|Immunogen||A synthetic peptide made to a C-terminal portion of the rat Calnexin protein (between residues 550-591) [UniProt P35565]|
Target Details CalnexinProduct Details anti-Calnexin Antibody Application Details Handling References for anti-Calnexin antibody (ABIN151474) Images back to top
|Alternative Name||Calnexin (CANX Antibody Abstract)|
|Background||Gene Symbol: CANX|
|Molecular Weight||Theoretical MW: 97 kDa|
|Pathways||MAPK Signaling, Thyroid Hormone Synthesis|
Application DetailsProduct Details anti-Calnexin Antibody Target Details Calnexin Handling References for anti-Calnexin antibody (ABIN151474) Images back to top
|Application Notes||Western Blot 1:1000, Simple Western 1:50, Flow Cytometry, Immunohistochemistry 1:100, Immunocytochemistry/Immunofluorescence 1:100, Immunoprecipitation 1:100, Immunohistochemistry-Paraffin 1:100This Calnexin antibody is useful for Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Western Blot, and Immunohistochemistry-paraffin embedded sections. In ICC/IF, endoplasmic reticulum staining was observed in HeLa cells. In Western Blot, a band is seen approx. 90 kDa representing Calnexin. In IHC-P, staining was observed in the endoplasmic reticulum of mouse bladder tisSue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer ( pH 6.0) is recommended. In Simple Western only 10 - 15 μL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Protocol specific for Calnexin antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room
|Restrictions||For Research Use only|
HandlingProduct Details anti-Calnexin Antibody Target Details Calnexin Application Details References for anti-Calnexin antibody (ABIN151474) Images back to top
PBS and 30 % Glycerol
Buffer contains: 0.05 % Sodium Azide
|Precaution of Use||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Handling Advice||Avoid freeze-thaw cycles|
|Storage||4 °C,-20 °C|
|Storage Comment||Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.|
References for anti-Calnexin antibody (ABIN151474)Product Details anti-Calnexin Antibody Target Details Calnexin Application Details Handling Images back to top
|Product cited in:||
Ye, Zhang, Ancrum, Manevich, Townsend, Tew: "Glutathione S-Transferase P-Mediated Protein S-Glutathionylation of Resident Endoplasmic Reticulum Proteins Influences Sensitivity to Drug-Induced Unfolded Protein Response." in: Antioxidants & redox signaling, 2016
Kang, Tang, Schapiro, Loux, Livesey, Billiar, Wang, Van Houten, Lotze, Zeh: "The HMGB1/RAGE inflammatory pathway promotes pancreatic tumor growth by regulating mitochondrial bioenergetics." in: Oncogene, Vol. 33, Issue 5, pp. 567-77, 2014 (Sample species: Mouse (Murine)). Further details: Western Blotting
Xu, Xu, Brown, Rossi, Hurd, Inturrisi, Pasternak, Pan: "Stabilization of the μ-opioid receptor by truncated single transmembrane splice variants through a chaperone-like action." in: The Journal of biological chemistry, Vol. 288, Issue 29, pp. 21211-27, 2013 (Sample species: Hamster). Further details: Western Blotting
Ghosh, Vo, Twiss, Kretz, Jozwiak, Montgomery, Shavit: "Characterization of zebrafish von Willebrand factor reveals conservation of domain structure, multimerization, and intracellular storage." in: Advances in hematology, Vol. 2012, pp. 214209, 2012 (Sample species: Human). Further details: Immunocytochemistry,Immunofluorescence
ImagesProduct Details anti-Calnexin Antibody Target Details Calnexin Application Details Handling References for anti-Calnexin antibody (ABIN151474) back to top