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AB257349

Human ALKBH5 knockout HEK-293T cell lysate

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ALKBH5 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 429 bp deletion in exon1.
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Western blot - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)
  • WB

Lab

Western blot - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : ALKBH5 knockout HEK-293T cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab195377 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab195377 was shown to react with ALKBH5 in wild-type HEK-293T cells in western blot with loss of signal observed in ALKBH5 knockout cell line ab266762 (ALKBH5 knockout cell lysate ab257349). Wild-type and ALKBH5 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab195377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ALKBH5 antibody [EPR18958] (<a href='/en-us/products/primary-antibodies/alkbh5-antibody-epr18958-ab195377'>ab195377</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

ALKBH5 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human ALKBH5 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-alkbh5-knockout-hek-293t-cell-line-ab266762'>ab266762</a>)

Predicted band size: 44 kDa

Observed band size: 48 kDa

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Sanger Sequencing - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)
  • Sanger seq

Unknown

Sanger Sequencing - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)

Allele-2 : 429 bp deletion in exon1

Sanger Sequencing - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)
  • Sanger seq

Unknown

Sanger Sequencing - Human ALKBH5 knockout HEK-293T cell lysate (AB257349)

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 429 bp deletion in exon1.

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Complementary Products

Primary Antibodies

AB195377

Anti-ALKBH5 antibody [EPR18958]

RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (AB195377)

  • 4
  • 4
Cell Lines & Lysates

AB263298

Human PIP5K1A knockout HEK-293T cell lysate

Sanger Sequencing - Human PIP5K1A knockout HEK-293T cell lysate (AB263298)

  • 0
  • 0
Primary Antibodies

AB189491

Anti-Heme Oxygenase 1 antibody [EPR18161-128]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (AB189491)

  • 5
  • 2
Proteins & Peptides

AB271361

Recombinant Human ALKBH5 protein (Tagged)

Functional Studies - Recombinant Human ALKBH5 protein (Tagged) (AB271361)

  • 0
  • 0
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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?

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Properties and storage information

Gene name
ALKBH5
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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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'.

For licensing inquiries, please contact partnerships@abcam.com