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AB263259

Human MRPS28 knockout HEK-293T cell lysate

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MRPS28 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.

View Alternative Names

28S ribosomal protein S28, 28S ribosomal protein S35, HSPC007, Mitochondrial 28S ribosomal protein S35, Mitochondrial ribosomal protein S28, RT28_HUMAN, S28mt, S35mt, mitochondrial

5 Images
Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)
  • WB

Lab

Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)

Lane 1 : Wild-type Raji cell lysate 20 μg

Lane 2 : MS4A1 knockout Raji cell lysate 20 μg

Lane 3 : A549 cell lysate 20 μg

Lane 4 : HeLa cell lysate 20 μg

False colour image of Western blot : Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) staining, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279298 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji 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.

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-mouse-igg1-chimeric-ab279298'>ab279298</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

MS4A1 knockout Raji cell lysate at 20 µg

Lane 2:

Western blot - Human MS4A1 (CD20) knockout Raji cell line (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)
  • WB

Lab

Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : MS4A1 knockout Raji cell lysate 20 μg
Lane 3 : A549 cell lysate 20 μg
Lane 4 : HeLa cell lysate 20 μg

False colour image of Western blot : Anti-CD20 antibody [EP459Y] – Mouse IgG2a (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279299 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji 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.

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-mouse-igg2a-chimeric-ab279299'>ab279299</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

MS4A1 knockout Raji cell lysate at 20 µg

Lane 2:

Western blot - Human MS4A1 (CD20) knockout Raji cell line (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)
  • WB

Lab

Western blot - Human MRPS28 knockout HEK-293T cell lysate (AB263259)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : MS4A1 knockout Raji cell lysate 20 μg
Lane 3 : A549 cell lysate 20 μg
Lane 4 : HeLa cell lysate 20 μg
False colour image of Western blot : Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279300 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji 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-Rat IgG H&L (IRDye® 800CW) preabsorbed (ab253031) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-rat-igg2a-chimeric-ab279300'>ab279300</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

MS4A1 knockout Raji cell lysate at 20 µg

Lane 2:

Western blot - Human MS4A1 (CD20) knockout Raji cell line (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Sanger Sequencing - Human MRPS28 knockout HEK-293T cell lysate (AB263259)
  • Sanger seq

Unknown

Sanger Sequencing - Human MRPS28 knockout HEK-293T cell lysate (AB263259)

Allele-2 : Insertion of the selection cassette in exon1

Sanger Sequencing - Human MRPS28 knockout HEK-293T cell lysate (AB263259)
  • Sanger seq

Unknown

Sanger Sequencing - Human MRPS28 knockout HEK-293T cell lysate (AB263259)

Allele-1 : 1 bp insertion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.

Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab263259-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human MRPS28 knockout HEK293T cell lysate", "number":"AB263259-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB263259-CMP02", "productcode":"" } ] } } }

Properties and storage information

Gene name
MRPS28
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

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

MRPS28 also known as Mitochondrial Ribosomal Protein S28 is a protein that plays an important role in mitochondrial protein synthesis. It is a component of the small subunit of the mitochondrial ribosome which is important for the translation of mitochondrial mRNAs. This protein has a molecular mass of approximately 12 kDa and is mainly expressed in the mitochondria an organelle present in most eukaryotic cells. As a mitochondrial ribosomal protein MRPS28 is part of the machinery responsible for producing proteins that are essential for mitochondrial function and energy production.
Biological function summary

MRPS28 is involved in the assembly and stability of the mitochondrial ribosomal small subunit complex. It ensures the proper translation of 13 mitochondrial-encoded polypeptides which are components of the oxidative phosphorylation system. MRPS28 interacts with other mitochondrial ribosomal proteins to form a functional ribosome supporting mitochondrial gene expression and cellular energy metabolism. The presence of MRPS28 in this complex is necessary for the maintenance of mitochondrial translation fidelity and efficiency.

Pathways

MRPS28 plays a role in mitochondrial translation and oxidative phosphorylation pathways. It contributes to the synthesis of proteins that are integral to the electron transport chain an important part of energy production in cells. Proteins such as MRPS16 and MRPS18 are also involved within these same pathways working closely with MRPS28 to ensure efficient mitochondrial function. These pathways are essential to provide energy in the form of ATP supporting various cellular processes.

MRPS28 has connections to conditions associated with mitochondrial dysfunction such as mitochondrial myopathy and Leigh syndrome. Research has suggested that mutations affecting MRPS28 could impair mitochondrial translation leading to decreased energy production and resulting in these disorders. Through its role in these diseases MRPS28 is linked with other mitochondrial proteins like MRPL3 which when dysfunctional can also contribute to similar mitochondrial pathologies. Understanding the function and disease associations of MRPS28 may provide insights into potential therapeutic strategies for mitochondrial diseases.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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For full details, please see our Terms & Conditions

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