Calnexin Antibody - BSA Free

Western Blot: Calnexin Antibody [NB100-1965] - WB analysis of CANX in cell lysates as noted.

Western Blot: Calnexin Antibody [NB100-1965] - Subcellular localization of TM6SF2.Immunoaffinity isolation of ER and Golgi complex from mouse liver. ER and Golgi fractions were prepared from mouse liver microsomes by immunoaffinity chromatography as described under Experimental Procedures. Microsome membranes were dissolved in RIPA buffer, and equal volumes were separated on 10% SDS-PAGE and immunoblotting as described under Experimental Procedures. BiP, binding immunoglobulin protein; Gos28, Golgi SNAP receptor complex member 1; *, nonspecific band. Image collected and cropped by Citeab from the following publication (Inactivation of Tm6sf2, a Gene Defective in Fatty Liver Disease, Impairs Lipidation but Not Secretion of Very Low Density Lipoproteins. J Biol Chem (2016) licensed under a CC-BY license.

Immunohistochemistry: Calnexin Antibody [NB100-1965] - Calnexin immunostaining in the anterior (AI) and mid (MI) intestine of ctrl (control zebrafish), D.I.O. (diet-induced obesity zebrafish), D.I.O. flw 3,5-T2 (D.I.O. zebrafish followed by 3,5-T2), D.I.O. with 3,5-T2 (D.I.O. zebrafish treated with 3,5-T2). (A) AI of ctrl zebrafish (B) AI of D.I.O. (C) AI of D.I.O. flw 3,5-T2. (D) AI of D.I.O. with 3,5-T2. The arrows indicate calnexin immunoexpression in the enteroendocrine and goblet cells. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2076-2615/10/7/1131/htm#) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with (NB100-1965) at 1 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with (NB100-1965) at 1 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - Rat FR cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with Calnexin Antibody (NB100-1965) at 1 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: Calnexin Antibody - BSA Free [NB100-1965] - Whole cell lysate or exosome from MDA-MD-231 cells was loaded with 20 ug/lane. 10% SDS-PAGE. Calnexin Antibody (NB100-1965) was used for primary antibody: 1:3000, 4C, overnight. Image from verified customer review.

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on rat FR cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: Calnexin Antibody [NB100-1965] - A dilution of 1/2000 used in TBS with 0.05% Tween-20. Bands visualized by ECL using goat anti-rabbit HRP at 1:2000.

Immunohistochemistry: Calnexin Antibody [NB100-1965] - IHC analysis of CANX in mouse prostate using DAB with hematoxylin counterstain.

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on Daudi cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on NIH3T3 cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Simple Western: Calnexin Antibody [NB100-1965] - Simple Western lane view shows a specific band for Calnexin in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ch, Dr, Gp, Rb, Sh, Xp, Ze, Ca
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Calnexin Antibody - BSA Free Summary

• Immunogen: A synthetic peptide made to an internal region of the canine Calnexin protein (within residues 25-100). [Swiss-Prot P24643]
• Localization: Endoplasmic reticulum
• Marker: Endoplasmic Reticulum Membrane Marker
• Predicted Species: Canine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CANX
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1-5 ug/ml
  • Immunocytochemistry/Immunofluorescence 1-5 ug/ml
  • Immunohistochemistry 1:40
  • Immunohistochemistry-Paraffin 1:40
  • Immunoprecipitation 1:100
  • Simple Western 1:25
  • Western Blot 2 ug/ml
• Application Notes: This Calnexin antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry-paraffin embedded sections, Immunoprecipitation and Western Blot. In Western blot a band is observed approx. 97 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 97 kDa.
  Disclaimer note: 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.

Reactivity Notes

Quail. Predicted to react with dog based on 100% sequence homology.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Calnexin Antibody - BSA Free

• calnexin
• CNX
• IP90FLJ26570
• Major histocompatibility complex class I antigen-binding protein p88
• P90

Background

Calnexin is a chaperone protein that assists in protein folding and quality control. Calnexin retains unassembled or unfolded N-linked glycoproteins in the endoplasmic reticulum and ensures that only properly assembled and folded proteins proceed down the secretory pathway. Calnexin antibodies can be used as endoplasmic reticulum markers (ER markers) in immunostaining experiments

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Calnexin Antibody (NB100-1965)(39)

Publications using NB100-1965

• Zhu Y, Hu S, Pan X et al. Hepatocyte Sirtuin 6 Protects against Atherosclerosis and Steatohepatitis by Regulating Lipid Homeostasis Cells 2023-08-05 [PMID: 37566087] (WB)
• Chen H, Qi X, Faulkner RA et al. Regulated degradation of HMG CoA reductase requires conformational changes in sterol-sensing domain Nature communications 2022-07-25 [PMID: 35879350] (WB, Chinese Hamster)
• Xu Y, Hu S, Jadhav K et al. Hepatocytic Activating Transcription Factor 3 Protects Against Steatohepatitis Via Hepatocyte Nuclear Factor 4 alpha Diabetes 2021-09-02 [PMID: 34475098]
• De Bardi M Pharmacological tuning of microRNAs in FAP-derived Extracellular Vesicles by HDAC 2 inhibitors promotes regeneration and reduces fibrosis in dystrophic muscles bioRxiv 2020-01-01 (WB, Mouse)
• Corbel S, Sampath S, Schmedt C, Zhang Q COMPOSITIONS, METHODS, AND THERAPEUTIC USES RELATED TO FUSOGENIC PROTEIN MINION United States Patent Application 20190263899 2019-08-29
• Elsabrouty R, Jo Y, Hwang S et al. Type 1 polyisoprenoid diphosphate phosphatase modulates geranylgeranyl-mediated control of HMG CoA reductase and UBIAD1 eLife 2021-11-29 [PMID: 34842525]
• Sung SE, Seo MS, Kang KK et al. Isolation and Characterization of Extracellular Vesicle from Mesenchymal Stem Cells of the Epidural Fat of the Spine Asian Spine Journal 2022-04-30 [PMID: 34461688] (B/N, WB)
• Zar� M, Campodonico J, Cosentino N et al. Plasma Exosome Profile in ST-Elevation Myocardial Infarction Patients with and without Out-of-Hospital Cardiac Arrest International Journal of Molecular Sciences 2021-07-28 [PMID: 34360827] (WB)
• Goudsmit, C, da Veiga Leprevost, F, et al. Differences in Extracellular Vesicle Protein Cargo Are Dependent on Head and Neck Squamous Cell Carcinoma Cell of Origin and Human Papillomavirus Status. Cancers (Basel) [PMID: 34359613] (Simple Western, Human)
• Xu Y, Zhu Y, Hu S et al. Hepatocyte miR-34a is a key regulator in the development and progression of non-alcoholic fatty liver disease Molecular metabolism 2021-04-27 [PMID: 33930596] (WB, Mouse)
• Millan C, Prause L, Vallmajo-Martin Q et al. Extracellular Vesicles from 3D Engineered Microtissues Harbor Disease-Related Cargo Absent in EVs from 2D Cultures Advanced healthcare materials 2021-04-22 [PMID: 33890421]
• Xu Y, Li Y, Jadhav K, et al. Hepatocyte ATF3 protects against atherosclerosis by regulating HDL and bile acid metabolism Nature metabolism 2021-01-01 [PMID: 33462514]
• Xu Y, Zhu Y, Hu S et al. Hepatocyte Nuclear Factor 4? Prevents the Steatosis-to-NASH Progression by Regulating p53 and Bile Acid Signaling (in mice) Hepatology 2021-06-01 [PMID: 33098092] (WB)
• Qi X, Friedberg L, De Bose-Boyd R et al. Sterols in an intramolecular channel of Smoothened mediate Hedgehog signaling Nat. Chem. Biol. 2020-09-14 [PMID: 32929279] (WB, Human)
• Sandona M, Consalvi S, et al. HDAC inhibitors tune miRNAs in extracellular vesicles of dystrophic muscle-resident mesenchymal cells. EMBO Rep 2020-09-03 [PMID: 32754983] (WB, Mouse)
• Imperatore R, Tunisi L, Mavaro I et al. Immunohistochemical Analysis of Intestinal and Central Nervous System Morphology in an Obese Animal Model (Danio rerio) Treated with 3,5-T2: A Possible Farm Management Practice? Animals (Basel) 2020-07-03 [PMID: 32635261] (IF/IHC, Zebrafish)
• Xu Y, Zhu Y, Bawa F et al. HepatocyteSpecific Expression of Human Carboxylesterase 1 Attenuates Diet-Induced Steatohepatitis and Hyperlipidemia in Mice Hepatol Commun 2020-02-20 [PMID: 32258948] (WB, Mouse)
• Jo Y, Kim Ss, Garland K et Al. Enhanced ER-associated degradation of HMG CoA reductase causes embryonic lethality associated with Ubiad1 deficiency Elife 2020-03-02 [PMID: 32118581] (IB, Mouse)
• Musante L, Bontha Sv, La Salvia S et Al. Rigorous characterization of urinary extracellular vesicles (uEVs) in the low centrifugation pellet - a neglected source for uEVs Sci Rep 2020-02-28 [PMID: 32111925] (WB, WB, Human)
  
  Details:
  Antibody was used in Western Blot analysis to verify that subcellular contaminants were not interfering with detecting target bio-markers.
• Xu Y, Xu Y, Zhu Y et al. Macrophage miR-34a Is a Key Regulator of Cholesterol Efflux and Atherosclerosis Mol. Ther. 2019-09-12 [PMID: 31604677]
• Khalaj K, Miller JE, Lingegowda H et al. Extracellular vesicles from endometriosis patients are characterized by a unique miRNA-lncRNA signature JCI Insight 2019-09-19 [PMID: 31534048] (WB, Human)
• Jo Y, Hamilton JS, Hwang S et al. Schnyder corneal dystrophy-associated UBIAD1 inhibits ER-associated degradation of HMG CoA reductase in mice Elife 2019-02-20 [PMID: 30785396] (WB, Mouse)
• Tiedemann K, Sadvakassova G, Mikolajewicz N et al. Exosomal Release of L-Plastin by Breast Cancer Cells Facilitates Metastatic Bone Osteolysis Transl Oncol 2018-12-20 [PMID: 30583289] (WB, Human)
• Gao HN, Guo HY, Zhang H et al. Yak milk-derived exosomes promote proliferation of intestinal epithelial cells in hypoxic environment J. Dairy Sci. 2018-12-20 [PMID: 30580945] (WB, Bovine)
• Qi X, Schmiege P, Coutavas E et al. Two Patched molecules engage distinct sites on Hedgehog yielding a signaling-competent complex Science 2018-08-23 [PMID: 30139912] (WB, Human)
• Qi X, Schmiege P, Coutavas E et al. Structures of human Patched and its complex with native palmitoylated sonic hedgehog. Nature. 2018-07-11 [PMID: 29995851] (WB, Human)
• Borgna V, Villegas J, Burzio VA et al. Mitochondrial ASncmtRNA-1 and ASncmtRNA-2 as potent targets to inhibit tumor growth and metastasis in the RenCa murine renal adenocarcinoma model Oncotarget 2017-07-04 [PMID: 28620146] (WB, Mouse)
• Xu Y, Li F, Zalzala M et al. Farnesoid X receptor activation increases reverse cholesterol transport by modulating bile acid composition and cholesterol absorption in mice Hepatology 2016-06-30 [PMID: 27359351]
• Smagris E, Gilyard S, BasuRay S et al. Inactivation of Tm6sf2, a Gene Defective in Fatty Liver Disease, Impairs Lipidation but Not Secretion of Very Low Density Lipoproteins. J Biol Chem 2016-05-13 [PMID: 27013658]
• McFarlane MR, Cantoria MJ, Linden AG et al. Scap is required for sterol synthesis and crypt growth in intestinal mucosa J. Lipid Res. 2015-04-20 [PMID: 25896350] (WB, Mouse)
• Turner N, Abeysinghe P, Sadowski P, Mitchell M Cross-species proteomic and microRNA comparison of extracellular vesicles in human milk, cow's milk, and infant formula products: moving towards next generation infant formula products bioRxiv 2023-02-24 (WB, Human, Bovine)
• Li N, Subrahmanyan L, Smith E et al. Mutations in the Histone Modifier PRDM6 Are Associated with Isolated Nonsyndromic Patent Ductus Arteriosus Am J Hum Genet 2016-05-17 [PMID: 27181681] (ICC/IF, Human)
• Huynh N, VonMoss L, Smith D et al. Characterization of Regulatory Extracellular Vesicles from Osteoclasts. J Dent Res 2016-06-01 [PMID: 26908631]
• Riemersma M, Sandrock J, Boltje TJ et al. Disease mutations in CMP-sialic acid transporter SLC35A1 result in abnormal alpha-dystroglycan O-mannosylation, independent from sialic acid. Hum. Mol. Genet. 2014-12-30 [PMID: 25552652] (WB, Human)
• Adamo G, Santonicola P, Picciotto S et al. Microalgae as a novel biofactory for biocompatible and bioactive extracellular vesicles bioRxiv 2023-04-04 (ICC/IF, Human)
• Saeki T, Sato K, Ito S et al. Importance of uncharged polar residues and proline in the proximal two-thirds (Pro107-Ser128) of the highly conserved region of mouse ileal Na+-dependent bile acid transporter, Slc10a2, in transport activity and cellular expression. BMC Physiol 2013-02-04 [PMID: 23374508] (WB, Porcine)
• Yuen MY, Webb SE, Chan CM et al. Characterization of Ca(2+) signaling in the external yolk syncytial layer during the late blastula and early gastrula periods of zebrafish development Biochim Biophys Acta 2012-11-08 [PMID: 23142640] (ICC/IF, IF/IHC, Zebrafish)
• Johnson B, Jun D, DeBose-Boyd R Membrane Topology of UbiA Prenyltransferase Domain-Containing Protein-1 (UBIAD1), a Novel Regulator of Cholesterol Homeostasis bioRxiv 2023-03-06 (WB)
• Tian E, Hoffman MP, Ten Hagen KG. O-glycosylation modulates integrin and FGF signalling by influencing the secretion of basement membrane components. Nat Commun 3:869. 2012-05-29 [PMID: 22643896] (ICC/IF, Mouse)

Reviews for Calnexin Antibody (NB100-1965) (4)

Average Rating: 4.3

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Mouse.

reviewed by: Yongkang Yang WB Human 10/21/2022

Summary

• Application: Western Blot
• Sample Tested: MDA MB 231 whole cell lysates and conditioned medium
• Species: Human
• Lot: E-2

reviewed by: Lamberto Torralba IP Human 03/14/2022

Summary

• Application: Immunoprecipitation
• Sample Tested: B cell lymphoma
• Species: Human
• Lot: E

Comments

• Comments: ***Bio-Techne Response: Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.***

reviewed by: Lamberto Torralba WB Human 02/16/2022

Summary

• Application: Western Blot
• Sample Tested: 293t HEK and U2OS cells
• Species: Human
• Lot: E

Comments

• Comments: Lane 1: WCL of 5 x 10^6 HEK cells
  Lane 2: Membrane enrichment of 5 x 10^6 HEK cells
  Size is around 75 kDA. One clear and specific band.

reviewed by: Elizabeth Soberg WB Mouse 06/13/2018

Summary

• Application: Western Blot
• Sample Tested: RAW 264.7 mouse monocyte/macrophage cell line
• Species: Mouse
• Lot: C-3

Comments

• Comments: 20 ug of whole cell lysate and exosomal lysate were loaded.
  Primary was used @ 2 ug/mL.
  Donkey anti-rabbit-HRP secondary was used @ 1:5000.
  Entry level (ug sensitivity) ECL reagents were used.

Bioinformatics

• Gene Symbol: CANX
• Entrez:
  • 29144(Rat)
• Uniprot:
  • P24643(Canine)