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AB266517

Human RTN3 (RTN3/HAP) knockout HEK-293T cell line

Human RTN3 (RTN3/HAP) knockout HEK-293T cell line

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RTN3 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

ASYIP, NSP-like protein 2, NSP-like protein II, NSPL2, NSPLII, Neuroendocrine-specific protein-like 2, Neuroendocrine-specific protein-like II, RTN3_HUMAN, Reticulon-3

2 Images
Western blot - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)
  • WB

Lab

Western blot - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)

False colour image of Western blot : Anti-RTN3/HAP antibody staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab68328 was shown to bind specifically to RTN3/HAP. A band was observed at 24 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in RTN3 knockout cell line ab266517 (knockout cell lysate ab258647). To generate this image, wild-type and RTN3 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-RTN3/HAP antibody (<a href='/products/primary-antibodies/rtn3-hap-antibody-ab68328'>ab68328</a>) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human RTN3 (RTN3/HAP) knockout HEK-293T cell lysate (<a href='/products/cell-lysates/human-rtn3-rtn3-hap-knockout-hek-293t-cell-lysate-ab258647'>ab258647</a>) at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

U-87 MG cell lysate at 20 µg

Predicted band size: 113 kDa

Observed band size: 24 kDa

false

Sanger Sequencing - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)
  • Sanger seq

Unknown

Sanger Sequencing - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)

Homozygous : 1 bp insertion in exon 1

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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製品の詳細

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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製品内容

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出荷温度及び保存条件

遺伝子名
RTN3
遺伝子編集のタイプ
Knockout
遺伝子編集の方法
CRISPR technology
ノックアウト検証方法
Sanger Sequencing
接合型
Homozygous
出荷温度
Dry Ice
短期保存温度
-196°C
長期保存温度
-196°C
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取り扱い方法

初回取り扱いガイドライン

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

継代培養ガイドライン
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

RTN3 also known as reticulon-3 or HAP (Hydroxy Alkenal Protection protein) is a member of the reticulon family of proteins which have roles in various cellular processes. This protein has a mass of approximately 130 kDa and is expressed in several tissues with significantly higher levels in the brain and spinal cord. RTN3 localizes to the endoplasmic reticulum where it participates in maintaining the structure of the tubular network influencing intracellular transport and signaling.
Biological function summary

The RTN3/HAP protein can influence neurite outgrowth implicating it in the development and maintenance of the nervous system. This protein associates closely with other members of the reticulon family forming complexes important for its functional roles within the cell. RTN3/HAP has been implicated in mechanisms that protect neurons from stress especially related to endoplasmic reticulum stress by its participation in membrane remodeling and regulation of lipid metabolism.

Pathways

Data shows that RTN3/HAP interacts with components of the endoplasmic reticulum stress response pathway and lipid biosynthesis pathways. It works alongside proteins such as RTN4/Nogo contributing to the regulation of axon guidance and regeneration. These pathways are essential for maintaining cellular homeostasis and responding to physiological stress with RTN3/HAP helping adapt or protect cells under challenging conditions.

Researchers have found associations between RTN3/HAP and neurodegenerative diseases like Alzheimer's disease where it is involved in pathologic processes such as amyloid-beta peptide aggregation. Aberrant expression or mutation of RTN3/HAP can disrupt normal protein turnover linking it to the pathophysiology of such diseases along with related proteins like presenilin and amyloid precursor protein (APP). Understanding these associations offers insights into therapeutic targets for managing these disorders.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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製品プロトコール

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ターゲットの情報

See full target information RTN3
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組合せ製品

Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB68328

Anti-RTN3/HAP antibody

KO Validated

Western blot - Anti-RTN3/HAP antibody (AB68328)

  • 3
  • 1
Proteins & Peptides

AB153066

Recombinant Human RTN3/HAP protein

SDS-PAGE - Recombinant Human RTN3/HAP protein (AB153066)

  • 0
  • 0
Primary Antibodies

AB250023

Anti-Nogo antibody [EPR12266] - BSA and Azide free

RabMAb|Recombinant

Flow Cytometry (Intracellular) - Anti-Nogo antibody [EPR12266] - BSA and Azide free (AB250023)

  • 0
  • 0

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