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RTN3 Recombinant Rabbit Monoclonal Antibody [PSH0-40] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750618

RTN3 Recombinant Rabbit Monoclonal Antibody [PSH0-40] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

RTN3 Recombinant Rabbit Monoclonal Antibody [PSH0-40] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human RTN3 aa 1-500 / 1,032.

Species Reactivity

Human, Rat

Validated Applications

WB, IF-Cell, FC

Target Molecular Weight

Predicted band size: 113 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, U87-MG cell lysate, A549 cell lysate, SH-SY5Y cell lysate, PC-12 cell lysate, L6 cell lysate, human cerebellum tissue lysate, PC-12, HeLa.

Conjugation

unconjugated

Clone Number

PSH0-40

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:50

  • FC

  • 1:500-1:1,000

Target

Function

May be involved in membrane trafficking in the early secretory pathway. Inhibits BACE1 activity and amyloid precursor protein processing. May induce caspase-8 cascade and apoptosis. May favor BCL2 translocation to the mitochondria upon endoplasmic reticulum stress. Induces the formation of endoplasmic reticulum tubules. Acts also as an inflammation-resolving regulator by interacting with both TRIM25 and RIG-I/DDX58, subsequently impairing DDX58 'Lys-63'-linked polyubiquitination leading to IRF3 and NF-kappa-B inhibition. This gene belongs to the reticulon family of highly conserved genes that are preferentially expressed in neuroendocrine tissues. This family of proteins interact with, and modulate the activity of beta-amyloid converting enzyme 1 (BACE1), and the production of amyloid-beta. An increase in the expression of any reticulon protein substantially reduces the production of amyloid-beta, suggesting that reticulon proteins are negative modulators of BACE1 in cells. Alternatively spliced transcript variants encoding different isoforms have been found for this gene, and pseudogenes of this gene are located on chromosomes 4 and 12.

Background References

1. Yang Z et al. RTN3 inhibits RIG-I-mediated antiviral responses by impairing TRIM25-mediated K63-linked polyubiquitination. Elife. 2021 Jul

2. Qian C et al. RTN3 - ASC interaction: The potential mechanism behind diabetes-induced cortical neuritic dystrophy. Neurosci Lett. 2022 Jan

Subcellular Location

Endoplasmic reticulum membrane, Golgi apparatus membrane.

UNIPROT

SwissProt: O95197 Human

SwissProt: Q6RJR6 Rat

Synonyms

ASYIP antibody

Neuroendocrine specific protein like 2 antibody

Neuroendocrine-specific protein-like 2 antibody

Neuroendocrine-specific protein-like II antibody

NSP like protein II antibody

NSP-like protein 2 antibody

NSP-like protein II antibody

NSPL2 antibody

NSPLII antibody

Reticulon 3 antibody

Expand

Images

  • Western blot analysis of RTN3 on different lysates with Rabbit anti-RTN3 antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (32 µg/Lane)<br />Lane 2: HEK-293 cell lysate (33 µg/Lane)<br />Lane 3: U87-MG cell lysate (30 µg/Lane)<br />Lane 4: A549 cell lysate (26 µg/Lane)<br />Lane 5: SH-SY5Y cell lysate (18 µg/Lane)<br />Lane 6: PC-12 cell lysate (23 µg/Lane)<br />Lane 7: L6 cell lysate (14 µg/Lane)<br />Lane 8: Human cerebellum tissue lysate (40 µg/Lane)<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 113 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of RTN3 on different lysates with Rabbit anti-RTN3 antibody (HA750618) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (32 µg/Lane)
    Lane 2: HEK-293 cell lysate (33 µg/Lane)
    Lane 3: U87-MG cell lysate (30 µg/Lane)
    Lane 4: A549 cell lysate (26 µg/Lane)
    Lane 5: SH-SY5Y cell lysate (18 µg/Lane)
    Lane 6: PC-12 cell lysate (23 µg/Lane)
    Lane 7: L6 cell lysate (14 µg/Lane)
    Lane 8: Human cerebellum tissue lysate (40 µg/Lane)

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 113 kDa
    Observed band size: 100 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750618) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of RTN3 on different lysates with Rabbit anti-RTN3 antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-RTN3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 113 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of RTN3 on different lysates with Rabbit anti-RTN3 antibody (HA750618) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-RTN3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 113 kDa
    Observed band size: 100 kDa

    Exposure time: 3 minutes; ECL: K1802;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750618) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of PC-12 cells labeling RTN3 with Rabbit anti-RTN3 antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RTN3 antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling RTN3 with Rabbit anti-RTN3 antibody (HA750618) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RTN3 antibody (HA750618) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling RTN3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750618" style="font-weight: bold;text-decoration: underline;">HA750618</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling RTN3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750618, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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