Anti-Calnexin antibody [6F12BE10] (ab112995)
![Western blot - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-260578-anti-calnexin-antibody-6f12be10-western-blot.jpg "Western blot - Anti-Calnexin antibody [6F12BE10] (ab112995)")
Key features and details
- Mouse monoclonal [6F12BE10] to Calnexin
- Suitable for: WB, IP, Flow Cyt, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG2b
Overview
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Product name
Anti-Calnexin antibody [6F12BE10]
See all Calnexin primary antibodies -
Description
Mouse monoclonal [6F12BE10] to Calnexin -
Host species
Mouse -
Specificity
Shotgun immunization of human HeLa cell lysates into mice. Targets were determined by mass spectrometry and validated by WB, ICC, ELISA pair and other techniques. -
Tested applications
Suitable for: WB, IP, Flow Cyt, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Human
Does not react with: Mouse -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa cells; HL60 cells and human fibroblasts.
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General notes
This monoclonal antibody to calnexin has been knockout validated in WB and ICC/IF. The expected signal was observed in wild type cells and the signal was not seen in knockout cells.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. -
Storage buffer
pH: 7.5
Preservative: 0.02% Sodium azide
Constituent: 99.98% HEPES buffered saline -
Concentration information loading... -
Purity
Proprietary Purification -
Purification notes
ab112995 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. Monoclonal purity was near homogeneity as judged by SDS-PAGE (>95%). -
Clonality
Monoclonal -
Clone number
6F12BE10 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab112995 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 |
|---|---|---|
| WB | (1) |
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
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| IP |
Use at an assay dependent concentration.
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| Flow Cyt |
Use a concentration of 1 µg/ml.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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| ICC/IF |
Use a concentration of 0.5 µg/ml.
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| IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa. |
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IP
Use at an assay dependent concentration. |
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Flow Cyt
Use a concentration of 1 µg/ml. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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ICC/IF
Use a concentration of 0.5 µg/ml. |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval 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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Western blot - Anti-Calnexin antibody [6F12BE10] (ab112995)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Calnexin knockout HAP1 cell lysate (20 µg)
Lane 3: THP1 cell lysate (20 µg)
Lane 4: Raw264.7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab112995 observed at 80 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab112995 was shown to specifically react with Calnexin. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab112995 at a concentration of 1 µg/mL and ab181602 (loading control to GAPDH) diluted to 1/1000 were incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
![Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-281490-anti-calnexin-antibody-6f12be10-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [6F12BE10] (ab112995)ab112995 staining Calnexin (shown in green) in wild-type HAP1 cells (top panel) and CANX knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 min and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab112995 at 0.5 μg/ml and ab202272 at 1/250 dilution (alpha tubulin shown in red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) 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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![Immunoprecipitation - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-125713-anti-calnexin-antibody-6f12be10-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Calnexin antibody [6F12BE10] (ab112995)Immunoprecipitation with ab112995.
Immunoprecipitation of Calnexin - ER membrane marker from HeLa cell lysate. The protein band runs around 90 kDa (predicted 68kDa) in tris-glycine SDS-PAGE. The identity of this protein was confirmed by mass spectrometry. This gel was stained with colloidal Coomassie blue G. -
![Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-125712-anti-calnexin-antibody-6f12be10-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [6F12BE10] (ab112995)ab112995 at 5µg/ml staining Calnexin - ER membrane marker in Human fibroblasts cells by Immunocytochemistry (4% paraformaldehyde fixed and 0.1% Triton X-100 permeabilized) followed by Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h (green).
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![Flow Cytometry - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-125711-anti-calnexin-antibody-6f12be10-flow-cytometry.jpg)
Flow Cytometry - Anti-Calnexin antibody [6F12BE10] (ab112995)ab112995 at 1µg/ml staining Calnexin - ER membrane marker in HL60 cells fixed with MeOH by Flow Cytometry (blue). Isotype control antibody (red). -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [6F12BE10] (ab112995)](https://www.abcam.com/ps/products/112/ab112995/Images/ab112995-125710-anti-calnexin-antibody-6f12be10-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [6F12BE10] (ab112995)IHC image of ab112995 staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab112995, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab112995 has been referenced in 10 publications.
- Ke Y et al. Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90. Stem Cell Res Ther 12:21 (2021). PubMed: 33413616
- Kweon HJ et al. NACHO Engages N-Glycosylation ER Chaperone Pathways for a7 Nicotinic Receptor Assembly. Cell Rep 32:108025 (2020). PubMed: 32783947
- Chen Y et al. MSC-Secreted Exosomal H19 Promotes Trophoblast Cell Invasion and Migration by Downregulating let-7b and Upregulating FOXO1. Mol Ther Nucleic Acids 19:1237-1249 (2020). PubMed: 32069774
- Wang J et al. Tracking extracellular vesicle phenotypic changes enables treatment monitoring in melanoma. Sci Adv 6:eaax3223 (2020). PubMed: 32133394
- Dafinca R et al. Impairment of Mitochondrial Calcium Buffering Links Mutations in C9ORF72 and TARDBP in iPS-Derived Motor Neurons from Patients with ALS/FTD. Stem Cell Reports 14:892-908 (2020). PubMed: 32330447
- Cook GM et al. Regulation of nerve growth and patterning by cell surface protein disulphide isomerase. Elife 9:N/A (2020). PubMed: 32452761
- Wang L et al. FAM92A1 is a BAR domain protein required for mitochondrial ultrastructure and function. J Cell Biol 218:97-111 (2019). PubMed: 30404948
- Nevzorov I et al. Myosin-1C uses a novel phosphoinositide-dependent pathway for nuclear localization. EMBO Rep 19:290-304 (2018). PubMed: 29330316
- Puri C et al. The RAB11A-Positive Compartment Is a Primary Platform for Autophagosome Assembly Mediated by WIPI2 Recognition of PI3P-RAB11A. Dev Cell 45:114-131.e8 (2018). ICC/IF . PubMed: 29634932
- Alpini C et al. Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques. Auto Immun Highlights 3:79-85 (2012). WB . PubMed: 26000130