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AB266765

Human ADRM1 (ARM-1) knockout HEK-293T cell line

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ADRM1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 16 bp deletion in exon 2.

View Alternative Names

110 kDa cell membrane glycoprotein, ADRM1_HUMAN, ARM-1, Adhesion-regulating molecule 1, Gp110, M(r) 110,000 surface antigen, Proteasomal ubiquitin receptor ADRM1, Proteasome regulatory particle non-ATPase 13, Regulatory particle non ATPase 13, Rpn13, Rpn13 homolog, hRpn13

4 Images
Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)
  • WB

Lab

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)

Lanes 1-4 : Merged signal (red and green). Green - ab157218 observed at 42 kDa. Red - loading control ab8245 observed at 36 kDa.

ab157218 Anti-ADRM1/ARM-1 antibody [EPR11450(B)] was shown to specifically react with ADRM1/ARM-1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266765 (knockout cell lysate ab257816) was used. Wild-type and ADRM1/ARM-1 knockout samples were subjected to SDS-PAGE. ab157218 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ADRM1/ARM-1 antibody [EPR11450(B)] (<a href='/en-us/products/primary-antibodies/adrm1-arm-1-antibody-epr11450b-ab157218'>ab157218</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ADRM1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (ab266765)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)
  • WB

Lab

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)

Lanes 1-4 : Merged signal (red and green). Green - ab157185 observed at 42 kDa. Red - loading control ab8245 observed at 36 kDa.

ab157185 Anti-ADRM1/ARM-1 antibody [EPR11449(B)] was shown to specifically react with ADRM1/ARM-1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266765 (knockout cell lysate ab257816) was used. Wild-type and ADRM1/ARM-1 knockout samples were subjected to SDS-PAGE. ab157185 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ADRM1/ARM-1 antibody [EPR11449(B)] (<a href='/en-us/products/primary-antibodies/adrm1-arm-1-antibody-epr11449b-ab157185'>ab157185</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ADRM1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (ab266765)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Sanger Sequencing - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)
  • Sanger seq

Unknown

Sanger Sequencing - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)

Homozygous : 16 bp deletion in exon 2

Cell Culture - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)
  • Cell Culture

Unknown

Cell Culture - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)

Representative images of ADRM1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 16 bp deletion in exon 2

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Reactivity data

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Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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

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What's included?

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Properties and storage information

Gene name
ADRM1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

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.

Subculture guidelines
  • 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.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

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

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Supplementary information

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

ADRM1 known also as ARM-1 or arm protein is an essential component of the proteasome system playing an important role in ubiquitin-proteasome pathway. Its molecular mass is about 42-46 kDa. ADRM1 protein is expressed across various tissues with significant presence in tissues requiring active protein degradation such as liver muscle and neurons. This protein interacts with other subunits in large protein complexes contributing to the regulation of protein turnover.
Biological function summary

The ADRM1/ARM-1 protein binds ubiquitinated substrates and shuttles them to the 26S proteasome for degradation. It acts as a receptor on the regulatory particle of the proteasome complex where it recruits deubiquitinating enzymes. This positions ADRM1 as an integral part of the protein degradation machinery ensuring removal of damaged misfolded or unnecessary proteins and maintaining cellular protein homeostasis.

Pathways

ADRM1 plays a significant role within the ubiquitin-proteasome pathway which is important for protein catabolism. It partners with regulatory proteins like Rpn13 to mediate the recognition and processing of polyubiquitinated substrates. The efficient functioning of this pathway highlights its importance in regulating the cell cycle and various signaling pathways including NF-kB signaling which affects immune response and apoptosis.

Dysfunction of ADRM1 can lead to pathological consequences. Its aberrant activity links to cancer particularly in solid tumors due to its role in controlling the degradation of cell cycle proteins. Additionally alterations in ADRM1 functionality can affect Alzheimer's disease as proper protein turnover is important in preventing accumulation of protein aggregates. Interactions with proteins like USP14 and UCHL5 further illustrate ADRM1's involvement in these pathologies marking it as a potential therapeutic target.
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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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Product protocols

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Target data

See full target information ADRM1
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