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AB256986

Human MCL1 knockout HEK-293T cell lysate

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MCL1 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 deletion in exon 1 and 7 bp insertion in exon 1.
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Western blot - Human MCL1 knockout HEK-293T cell lysate (AB256986)
  • WB

Lab

Western blot - Human MCL1 knockout HEK-293T cell lysate (AB256986)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : MCL1 knockout HEK293T cell lysate (20 ug)
Lane 3 : Ramos cell lysate (20 ug)

ab32087 was shown to specifically react with MCL1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266838 (knockout cell lysate ab256986) was used. Wild-type and MCL1 knockout samples were subjected to SDS-PAGE. ab32087 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4oC 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-MCL1 antibody [Y37] (<a href='/en-us/products/primary-antibodies/mcl1-antibody-y37-ab32087'>ab32087</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MCL1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MCL1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mcl1-knockout-hek-293t-cell-line-ab266838'>ab266838</a>)

Lane 3:

Ramos 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: 37 kDa

Observed band size: 37 kDa

false

Sanger Sequencing - Human MCL1 knockout HEK-293T cell lysate (AB256986)
  • Sanger seq

Unknown

Sanger Sequencing - Human MCL1 knockout HEK-293T cell lysate (AB256986)

Allele-1 : 1 bp deletion in exon 1

Sanger Sequencing - Human MCL1 knockout HEK-293T cell lysate (AB256986)
  • Sanger seq

Unknown

Sanger Sequencing - Human MCL1 knockout HEK-293T cell lysate (AB256986)

Allele-2 : 7 bp insertion in exon 1

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 deletion in exon 1 and 7 bp insertion in exon 1.

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Complementary Products

Cell Lines & Lysates

AB266838

Human MCL1 knockout HEK-293T cell line

Advanced Validation

Western blot - Human MCL1 knockout HEK-293T cell line (AB266838)

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Primary Antibodies

AB32087

Anti-MCL1 antibody [Y37]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-MCL1 antibody [Y37] (AB32087)

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Proteins & Peptides

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Recombinant human Cleaved Caspase-3 protein (Active)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

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Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

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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
MCL1
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.

MCL1 also known as myeloid cell leukemia sequence 1 is an anti-apoptotic member of the BCL-2 family. This protein plays an important role in regulating cell survival by inhibiting apoptosis. MCL1 has a molecular weight of approximately 37 kDa. It is primarily expressed in the mitochondria and can be found in various tissues including heart liver placenta and kidneys.
Biological function summary

MCL1 protein functions to maintain cell viability by counteracting pro-apoptotic stimuli. It interacts within the cell as part of a complex network involving other BCL-2 family proteins. MCL1 binds and sequesters BH3-only proteins and Bax therefore preventing them from initiating the apoptotic cascade. This interaction helps cells survive under stress conditions by temporarily inhibiting apoptosis.

Pathways

MCL1 plays a significant role in the intrinsic (mitochondrial) apoptosis pathway and the cell signaling pathways that facilitate cell survival and proliferation. In the mitochondrial pathway it works closely with other BCL-2 family members like BCL-xL and BCL-2 itself to control the permeability of the mitochondrial outer membrane and thereby influence cytochrome c release. The PI3K/AKT pathway can also regulate MCL1 leading to increases in its expression and stability which promotes cell survival.

Researchers have linked alterations in MCL1 expression to various cancers including leukemia and lymphoma. Overexpression of MCL1 can contribute to chemoresistance in cancer cells by preventing apoptosis. This protein's function is also connected to cardiomyopathies where altered MCL1 expression in cardiac tissue can influence cell death and survival outcomes. BCL-2 and BCL-xL closely related proteins also implicate in these conditions highlighting the central role of the apoptotic regulatory network in disease development.
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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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