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AB257378

Human CAPG (MCP) knockout HeLa cell lysate

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CAPG 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 exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.

View Alternative Names

Actin regulatory protein CAP-G, Macrophage-capping protein, MCP, CAPG, AFCP

5 Images
Western blot - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)
  • WB

Lab

Western blot - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : CAPG knockout HeLa cell lysate (20 ug)
Lane 3 : THP-1 cell lysate (20 ug)
Lane 4 : U-937 cell lysate (20 ug)

ab181092 was shown to specifically react with Actin Regulatory Protein CAPG/MCP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265766 (knockout cell lysate ab257378) was used. Wild-type and Actin Regulatory Protein CAPG/MCP knockout samples were subjected to SDS-PAGE. ab181092 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-Actin Regulatory Protein CAPG/MCP antibody [EPR13194] (<a href='/en-us/products/primary-antibodies/actin-regulatory-protein-capg-mcp-antibody-epr13194-ab181092'>ab181092</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAPG knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAPG (MCP) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-capg-mcp-knockout-hela-cell-line-ab265766'>ab265766</a>)

Lane 3:

THP-1 cell lysate at 20 µg

Lane 4:

U-937 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: 38 kDa

Observed band size: 48 kDa

false

Western blot - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)
  • WB

Lab

Western blot - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : CAPG knockout HeLa cell lysate (20 ug)
Lane 3 : THP-1 cell lysate (20 ug)
Lane 4 : U-937 cell lysate (20 ug)

ab181095 was shown to specifically react with Actin Regulatory Protein CAPG/MCP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265766 (knockout cell lysate ab257378) was used. Wild-type and Actin Regulatory Protein CAPG/MCP knockout samples were subjected to SDS-PAGE. ab181095 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-Actin Regulatory Protein CAPG/MCP antibody [EPR13194-34] (<a href='/en-us/products/primary-antibodies/actin-regulatory-protein-capg-mcp-antibody-epr13194-34-ab181095'>ab181095</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAPG knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAPG (MCP) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-capg-mcp-knockout-hela-cell-line-ab265766'>ab265766</a>)

Lane 3:

THP-1 cell lysate at 20 µg

Lane 4:

U-937 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: 38 kDa

Observed band size: 48 kDa

false

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)

Allele-1 : 2 bp deletion in exon 2

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)

Allele-3 : Insertion of the selection cassette in exon 2

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPG (MCP) knockout HeLa cell lysate (AB257378)

Allele-2 : 1 bp insertion in exon 2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.

Disease

Adenocarcinoma

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Cell Lines & Lysates

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Advanced Validation

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Primary Antibodies

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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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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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257378-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human CAPG knockout HeLa cell lysate", "number":"AB257378-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257378-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
CAPG
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
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Supplementary information

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

Actin Regulatory Protein CAPG also known as MCP is an important cytoskeletal protein involved in actin filament dynamics. Other common names include CAPG protein and capping proteins. CAPG binds to the barbed ends of actin filaments modulating their length and polymerization. This protein has a molecular weight of approximately 39 kDa. It is expressed in various tissues with significant expression in HeLa cells which are often used in studies related to actin regulation.
Biological function summary

This actin regulatory protein serves as an important player in the remodeling of the actin cytoskeleton. CAPG functions by capping the fast-growing ends of actin filaments which influences cell motility and structure. It is not known to be part of a larger protein complex but interacts closely with other actin-modulating proteins. MCP's regulation of cytoskeletal dynamics allows cells to respond swiftly to environmental changes assisting cellular processes like migration and secretion.

Pathways

CAPG is intricately involved in pathways controlling cell motility and structure including the cytoskeletal rearrangement pathway. It works alongside proteins such as cofilin and Arp2/3 complex which also regulate actin dynamics. These proteins together facilitate processes like cell division and migration making their interactions essential for proper cellular function.

CAPG's regulation is pertinent in cancer metastasis particularly in its relation to HeLa cells used in cancer studies. It affects tumor cell invasiveness by altering the cytoskeleton connecting it to processes involving other regulatory proteins like Rac1. Additionally CAPG plays a role in neurodegenerative disorders by impacting neuronal cell motility where interactions with actin-modulating proteins are important. Understanding CAPG's role gives insight into potential interventions for 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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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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