HRD1 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-2526

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Hamster

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all HRD1 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of human HRD1 (within residues 300-400). [UniProt Q96PK3]

Reactivity Notes

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

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HRD1 Antibody - BSA Free

Western Blot: HRD1 AntibodyBSA Free [NB100-2526]

Western Blot: HRD1 AntibodyBSA Free [NB100-2526]

HRD1-Antibody-Western-Blot-NB100-2526-img0006.jpg
Immunocytochemistry/ Immunofluorescence: HRD1 Antibody - BSA Free [NB100-2526]

Immunocytochemistry/ Immunofluorescence: HRD1 Antibody - BSA Free [NB100-2526]

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 - BSA Free [NB100-2526]

Immunohistochemistry-Paraffin: HRD1 Antibody - BSA Free [NB100-2526]

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 AntibodyBSA Free [NB100-2526]

Western Blot: HRD1 AntibodyBSA Free [NB100-2526]

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.

Applications for HRD1 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:200 - 1:500

Immunohistochemistry-Paraffin

1:200 - 1:500. Use reported in scientific literature (PMID 24453213)

Immunoprecipitation

reported in scientific literature (PMID 35350536)

Knockout Validated

reported in scientific literature (PMID 26551274)

Western Blot

1:1000 - 1:2000
Application Notes
Heat samples at 50C for 30 minutes to detect HRD1 by Western blot.

Reviewed Applications

Read 3 reviews rated 5 using NB100-2526 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: HRD1

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.

Alternate Names

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

Entrez Gene IDs

84447 (Human); 74126 (Mouse)

Gene Symbol

SYVN1

UniProt

Q86TM6 (Human)

Additional HRD1 Products

Product Documents for HRD1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for HRD1 Antibody - BSA Free

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.

Citations for HRD1 Antibody - BSA Free

Customer Reviews for HRD1 Antibody - BSA Free (3)

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Customer Images

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  • HRD1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: RKO and AR42J human acinar pancreatic cancer cell line
    Species: Human
    Verified Customer | Posted 04/04/2020
    HRD1 Antibody - BSA Free NB100-2526
  • HRD1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T and MDA MB 231 cells
    Species: Human
    Verified Customer | Posted 11/06/2018
    HRD1 Antibody - BSA Free NB100-2526
  • HRD1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T and HePG2 cell lysate
    Species: Human
    Verified Customer | Posted 10/17/2018
    HRD1 Antibody - BSA Free NB100-2526

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Protocols

View specific protocols for HRD1 Antibody - BSA Free (NB100-2526):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen 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 (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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FAQs for HRD1 Antibody - BSA Free

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  • Q: What is the molecular weight for HRD1?

    A: The theoretical molecular weight for HRD1 is 67kDa.

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