HRD1 Antibody

Western Blot: HRD1 Antibody [NB100-2526] - Hrd1 is required for CDT intoxication. Co-immunoprecipitation of Derl2 and Hrd1. Derl2 was immunoprecipitated and samples were analyzed for Hrd1 by western blot. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.ppat.1004295) licensed under a CC-BY licence.

Immunocytochemistry/Immunofluorescence: HRD1 Antibody [NB100-2526] - HRD1 antibody was tested in Hek293 cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

Immunohistochemistry-Paraffin: HRD1 Antibody [NB100-2526] - FFPE tissue section of human tonsil using rabbit polyclonal HRD1 antibody at 2 ug/mL concentration (1:500 dilution). The primary antibody binding to SYVN1/HRD1 antigen in cells was detected using HRP conjugated anti-Rabbit secondary antibody with DAB reagent, and the sections were further counterstained with hematoxylin for labeling cellular nuclei. This HRD1 antibody generated an expected cytoplasmic staining of HRD1 with stronger signal around periphery of the nucleus signifying the staining of the endoplasmic reticulum.

Western Blot: HRD1 Antibody [NB100-2526] - Total protein from human HeLa, Hek293 and mouse 3T3 cell lines was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-HRD1 in block buffer and detected with an anti-rabbit HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent.

• Reactivity: Hu, Mu, Ha
• Applications: WB, ICC/IF, IHC, IHC-P, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

HRD1 Antibody Summary

• Immunogen: A synthetic peptide made to an internal portion of human HRD1 (within residues 300-400). [UniProt Q96PK3]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: SYVN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000 - 1:2000
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunohistochemistry 1:200 - 1:500
  • Immunohistochemistry-Paraffin 1:200 - 1:500
  • Knockout Validated
• Application Notes: Heat samples at 50C for 30 minutes to detect HRD1 by Western blot. Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 24453213). KO Validation was reported in scientific literature (PMID: 26551274).
• Control:

  HRD1 Overexpression Lysate

Reactivity Notes

Hamster reactivity reported in scientific literature (PMID: 25078082).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HRD1 Antibody

• E3 ubiquitin-protein ligase synoviolin
• EC 6.3.2
• HRD1EC 6.3.2.-
• KIAA1810Synovial apoptosis inhibitor 1
• MGC40372
• synovial apoptosis inhibitor 1, synoviolin

Background

HRD1 (synovial apoptosis inhibitor 1 or SYVN1) is a ubiquitin ligase whose expression is induced by UPR following ER stress and it provides protection to cells from apoptosis by inducing degradation of abnormally processed proteins that accumulate in ER. HRD1 acts as an E3 ubiquitin-protein ligase which accepts ubiquitin specifically from ER-associated UBC7 E2 ligase and transfers it to target proteins for promoting their degradation. It is a component of ER's quality control system called ERAD (ER-associated degradation) which is involved in ubiquitin-dependent degradation of misfolded ER proteins. HRD1 also facilitates the degradation of normal/naturally short-lived proteins such as SGK, and has been shown to protect neuronal cells from apoptosis triggered by polyglutamine-expanded huntingtin (HTT) or unfolded GPR37 by prooting their degradation. Moreover, HRD1 sequesters p53/TP53 in the cytoplasm and promotes its degradation, thereby negatively regulating its biological function in transcription, cell cycle regulation and apoptosis.

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 HRD1 Antibody (NB100-2526)(11)

Publications using NB100-2526

• Jiang LL, Zhu B, Zhao Y et al. Membralin deficiency dysregulates astrocytic glutamate homeostasis leading to ALS-like impairment J. Clin. Invest. May 21 2019 [PMID: 31112137] (IHC-P, Human, Mouse)
• Xie H, Tang CA, Song JH, Mancuso A. IRE1a RNase-dependent lipid homeostasis promotes survival in Myc-transformed cancers. J. Clin. Invest. 2018 Feb 26 [PMID: 29381485] (WB, Human)
• Hwang J, Walczak CP, Shaler TA et al. Characterization of protein complexes of the endoplasmic reticulum associated degradation E3 ubiquitin ligase Hrd1. J. Biol. Chem. Apr 14 2017 [PMID: 28411238] (WB, Human)
• Koenig Pa, Nicholls Pk, Schmidt Fi et al. The E2 Ubiquitin Conjugating Enzyme UBE2J1 is Required for Spermiogenesis in Mice. J. Biol. Chem. 2014 Oct 15 [PMID: 25320092] (WB, Mouse)
• Eshraghi A, Dixon SD, Tamilselvam B et al. Cytolethal Distending Toxins Require Components of the ER-Associated Degradation Pathway for Host Cell Entry PLoS Pathog. 2014 Jul 01 [PMID: 25078082]
• Sun S, Shi G, Han X et al. Sel1L is indispensable for mammalian endoplasmic reticulum-associated degradation, endoplasmic reticulum homeostasis, and survival. Proc. Natl. Acad. Sci. U.S.A. 2014 Feb 5 [PMID: 24453213] (WB, IHC-P, Mouse)
• Francisco AB, Singh R, Sha H et al. Haploid insufficiency of suppressor enhancer Lin12 1 like (SEL1L) predisposes mice to high fat diet-induced hyperglycemia. J Biol Chem;286(25):22275-82. 2011 Jun 24. [PMID: 21536682]
• Dougan SK, Hu CC, Paquet ME et al. Derlin-2-deficient mice reveal an essential role for protein dislocation in chondrocytes Mol Cell Biol 2011 Mar [PMID: 21220515] (WB, Mouse)
• Soundararajan R, Wang J, Melters D, Pearce D. Glucocorticoid-induced Leucine zipper 1 stimulates the epithelial sodium channel by regulating serum- and glucocorticoid-induced kinase 1 stability and subcellular localization. J Biol Chem 2010 Dec 17 [PMID: 20947508] (WB, Human)
• Sun S, Shi G, Sha H et al. IRE1 [alpha] is an endogenous substrate of endoplasmic-reticulum-associated degradation. Nature cell Biology. 2015 Nov 09 (WB, Mouse)
• Sun S, Shi G, Sha H et al. IRE1alpha is an endogenous substrate of endoplasmic-reticulum-associated degradation. Nat Cell Biol 2015 Dec [PMID: 26551274] (WB)

Reviews for HRD1 Antibody (NB100-2526) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 1 species: Human.

reviewed by: Anonymous WB Human 04/04/2020

Summary

• Application: Western Blot
• Sample Tested: RKO,AR42J human acinar pancreatic cancer cell line
• Species: Human
• Lot: A-2

reviewed by: Anonymous WB Human 11/06/2018

Summary

• Application: Western Blot
• Sample Tested: 293T,MDA MB 231 cells
• Species: Human
• Lot: A-2

reviewed by: Anonymous WB Human 10/17/2018

Summary

• Application: Western Blot
• Sample Tested: 293T and HePG2 cell lysate
• Species: Human
• Lot: A-2

FAQs for HRD1 Antibody (NB100-2526). (Showing 1 - 1 of 1 FAQs).

1. What is the molecular weight for HRD1?
   • The theoretical molecular weight for HRD1 is 67kDa.

Bioinformatics

• Gene Symbol: SYVN1
• Entrez:
  • 84447(Human)
  • 74126(Mouse)
• Uniprot:
  • Q86TM6(Human)