Recombinant Anti-Calnexin antibody [EPR3632] (ab92573)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3632] to Calnexin
- Suitable for: ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Calnexin antibody [EPR3632]
See all Calnexin primary antibodies -
Description
Rabbit monoclonal [EPR3632] to Calnexin -
Host species
Rabbit -
Specificity
Recognizes ER membrane, mitochondria and cis-Golgi -
Tested applications
Suitable for: ICC/IF, WB, IP, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Calnexin aa 1-100. The exact sequence is proprietary.
Database link: P27824 -
Positive control
- WB: HeLa, A431, SH-SY5Y, HEK-293T, MCF7, U-2 OS and HepG2 whole cell lysate (ab7900). IHC-P: Human tonsil tissue. ICC/IF: Wild-type HAP1 cells. IP: HeLa lysate.
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General notes
References regarding specificity:
Horner SM et al. Mitochondrial-associated endoplasmic reticulum membranes (MAM) form innate immune synapses and are targeted by hepatitis C virus. Proc Natl Acad Sci U S A 108:14590-5 (2011). PubMed: 21844353
Myhill N et al. The subcellular distribution of calnexin is mediated by PACS-2. Mol Biol Cell 19:2777-88 (2008). PubMed: 18417615
Yoshimura SI et al. Direct targeting of cis-Golgi matrix proteins to the Golgi apparatus. J Cell Sci 114:4105-15 (2001). PubMed: 11739642
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3632 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92573 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/1000.
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WB |
1/20000 - 1/100000. Predicted molecular weight: 90 kDa.
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IP |
1/50.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ICC/IF
1/1000. |
WB
1/20000 - 1/100000. Predicted molecular weight: 90 kDa. |
IP
1/50. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins. -
Sequence similarities
Belongs to the calreticulin family. -
Cellular localization
Endoplasmic reticulum membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 821 Human
- Omim: 114217 Human
- SwissProt: P27824 Human
- Unigene: 567968 Human
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Alternative names
- Calnexin antibody
- CALX_HUMAN antibody
- CANX antibody
see all
Images
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All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CANX knockout HEK-293T cell lysate
Lane 3 : U-2 OS cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 90 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab92573 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92573 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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ab92573 staining Calnexin in wild-type HAP1 cells (top panel) and CANX knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92573 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : Calnexin knockout HAP1 cell lysate
Lane 3 : THP-1 cell lysate
Lane 4 : RAW 264.7 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 90 kDaLanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab92573 was shown to specifically react with Calnexin when Calnexin knockout samples were used. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab92573 and ab8245 (loading control to GAPDH) were diluted at 1/20,000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging. -
All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A431 cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : HepG2 cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : standard HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 90 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] (ab92573)
Immunohistochemical analysis of paraffin embedded Human tonsil tissue using ab92573 at a 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Calnexin was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with 92573 at 1/100 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab92573 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab92573 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-Calnexin antibody [EPR3632] (ab92573)
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : Calnexin knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 90 kDaLanes 1 - 2: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab92573 and a competitor's rabbit polyclonal antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (38)
ab92573 has been referenced in 38 publications.
- Scott TA et al. Engineered extracellular vesicles directed to the spike protein inhibit SARS-CoV-2. Mol Ther Methods Clin Dev 24:355-366 (2022). PubMed: 35127966
- Tian J et al. Extracellular vesicles from bone marrow stromal cells reduce the impact of stroke on glial cell activation and blood brain-barrier permeability via a putative miR-124/PRX1 signalling pathway. Eur J Neurosci 56:3786-3805 (2022). PubMed: 35441400
- Nakano K et al. Bioactive Evaluation of Ursane-Type Pentacyclic Triterpenoids: β-Boswellic Acid Interferes with the Glycosylation and Transport of Intercellular Adhesion Molecule-1 in Human Lung Adenocarcinoma A549 Cells. Molecules 27:N/A (2022). PubMed: 35630550
- Keremu A et al. Extracellular vesicles from bone mesenchymal stem cells transport microRNA-206 into osteosarcoma cells and target NRSN2 to block the ERK1/2-Bcl-xL signaling pathway. Eur J Histochem 66:N/A (2022). PubMed: 35730574
- Desoutter A et al. Enamel and dentin in Enamel renal syndrome: A confocal Raman microscopy view. Front Physiol 13:957110 (2022). PubMed: 36091358