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AB256946

Human HTT (Huntingtin) knockout HeLa cell lysate

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HTT KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon21.

View Alternative Names

AI256365, C430023I11Rik, HD protein, HDH, HD_HUMAN, HUNTINGTON CHOREA, Huntingtin, Huntington disease protein, Huntington's disease protein homolog, IT 15, OTTMUSP00000026909, ZHD

3 Images
Western blot - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)
  • WB

Lab

Western blot - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : HTT knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab45169 observed at 348 kDa. Red - loading control ab7291 observed at 50 kDa.
ab45169 Anti-Huntingtin antibody [EP867Y] was shown to specifically react with Huntingtin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265976 (knockout cell lysate ab256946) was used. Wild-type and Huntingtin knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab45169 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 10000 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-Huntingtin antibody [EP867Y] (<a href='/en-us/products/primary-antibodies/huntingtin-antibody-ep867y-ab45169'>ab45169</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HTT knockout HeLa cell lysate at 20 µg

Predicted band size: 348 kDa

Observed band size: 348 kDa

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Western blot - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)
  • WB

Lab

Western blot - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : HTT knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab109115 observed at 348 kDa. Red - loading control ab7291 observed at 50 kDa.
ab109115 Anti-Huntingtin antibody [EPR5526] was shown to specifically react with Huntingtin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265976 (knockout cell lysate ab256946) was used. Wild-type and Huntingtin knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109115 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 10000 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-Huntingtin antibody [EPR5526] (<a href='/en-us/products/primary-antibodies/huntingtin-antibody-epr5526-ab109115'>ab109115</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HTT knockout HeLa cell lysate at 20 µg

Predicted band size: 348 kDa,43 kDa,52 kDa,55 kDa,56 kDa

Observed band size: 348 kDa

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Sanger Sequencing - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)
  • Sanger seq

Unknown

Sanger Sequencing - Human HTT (Huntingtin) knockout HeLa cell lysate (AB256946)

Homozygous : 1 bp deletion in exon21

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon21.

Disease

Adenocarcinoma

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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>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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 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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256946-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human HTT knockout HeLa cell lysate", "number":"AB256946-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256946-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
HTT
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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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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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Complementary Products

Primary Antibodies

AB109115

Anti-Huntingtin antibody [EPR5526]

KO Validated|RabMAb|Recombinant|20ul selling size

Western blot - Anti-Huntingtin antibody [EPR5526] (AB109115)

  • 5
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Cell Lines & Lysates

AB265976

Human HTT (Huntingtin) knockout HeLa cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human HTT (Huntingtin) knockout HeLa cell line (AB265976)

  • 0
  • 0
Primary Antibodies

AB129002

Anti-Vinculin antibody [EPR8185] - Loading Control

20ul selling size|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

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  • 21
Proteins & Peptides

AB112300

Recombinant Human Huntingtin protein (Tagged)

SDS-PAGE - Recombinant Human Huntingtin protein (Tagged) (AB112300)

  • 0
  • 0

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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