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AB257274

Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate

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SETD2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 3 and 4 bp deletion in exon 3.

View Alternative Names

Huntingtin-interacting protein B, p231HBP, HIF-1, HSPC069, Huntingtin yeast partner B, HYPB, Lysine N-methyltransferase 3A, HIF1, SETD2, Histone-lysine N-methyltransferase SETD2, Huntingtin-interacting protein 1, KMT3A, SET2, SET domain-containing protein 2, hSET2, HIP-1, Protein-lysine N-methyltransferase SETD2, KIAA1732

5 Images
Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)
  • WB

Unknown

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)

Lane 1 : Wild-type HEK293T cell lysate (20 μg)

Lane 2 : SETD2 knockout HEK293T cell lysate (20 μg)

Lane 3 : HepG2 cell lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab31358 observed at 288 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab31358 Anti-KMT3A / HYPB / HIF-1 antibody was shown to specifically react with KMT3A / HYPB / HIF-1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266690 (knockout cell lysate ab257274) was used. Wild-type and KMT3A / HYPB / HIF-1 knockout samples were subjected to SDS-PAGE. ab31358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-ab31358'>ab31358</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

SETD2 knockout HEK293T cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Predicted band size: 288 kDa

Observed band size: 288 kDa

false

Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)
  • Sanger seq

Unknown

Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)

Allele-2 : 4 bp deletion in exon 3

Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)
  • Sanger seq

Unknown

Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)

Allele-1 : 11 bp deletion in exon 3

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)
  • WB

Lab

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)

*Lanes 1-3 : * Merged signal (red and green). Green – ab31358 observed at 288 kDa. Red - loading control, ab7291 observed at 50 kDa. ab31358 was shown to react with SETD2 in wild-type HEK293T cells in Western blot. Loss of signal was observed when knockout sample ab257274 was used. Wild-type and SETD2 knockout samples were subjected to SDS-PAGE. ab31358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-ab31358'>ab31358</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell at 20 µg

Lane 2:

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (ab257274) at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>)

Lanes 1 - 3:

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: 288 kDa

Observed band size: 288 kDa

false

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)
  • WB

Lab

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (AB257274)

*Lanes 1-3 : * Merged signal (red and green). Green – ab31358 observed at 288 kDa. Red - loading control, ab7291 observed at 50 kDa. ab31358 was shown to react with SETD2 in wild-type HEK293T cells in Western blot. Loss of signal was observed when knockout sample ab257274 was used. Wild-type and SETD2 knockout samples were subjected to SDS-PAGE. ab31358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-ab31358'>ab31358</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell at 20 µg

Lane 2:

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HEK-293T cell lysate (ab257274) at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Predicted band size: 288 kDa

Observed band size: 288 kDa

false

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, 11 bp deletion in exon 3 and 4 bp deletion in exon 3.

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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 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?

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Properties and storage information

Gene name
SETD2
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.

KMT3A also known as HYPB (Huntington's disease gene-related protein B) or HIF-1 is a histone methyltransferase enzyme that mechanically transfers methyl groups to histone H3 at lysine 4 and lysine 36. This activity important for chromatin modification influences transcriptional regulation. The molecular mass of KMT3A is approximately 130 kDa. This enzyme is expressed in various tissues such as the brain heart and skeletal muscle highlighting its extensive physiological role.
Biological function summary

KMT3A modifies chromatin structure to regulate gene expression. Its histone methylation function plays a critical role in the modulation of transcriptional activity ensuring that genes are turned on or off when necessary for cellular function. Additionally KMT3A forms part of complex protein networks that mediate gene expression changes in response to cellular signals. The precise regulation of this activity is vital for maintaining normal cellular function and responding to environmental cues.

Pathways

KMT3A engages in significant pathways such as the transcriptional regulation and epigenetic modification pathways. It interacts with various proteins including CoREST and LSD1 within these pathways to modulate gene expression and influence cellular differentiation. Through these interactions KMT3A ensures the correct genetic programs are executed aligning with developmental and homeostatic requirements.

KMT3A has been implicated in neurological conditions such as Huntington's disease. The abnormal regulation or mutation of this target can disrupt its activity and lead to improper chromatin modification which results in altered gene expression and contributes to disease progression. Furthermore KMT3A's interaction with the HIF-1α pathway also links it to cancer as dysregulation can aid in tumor growth and progression by affecting gene expression patterns associated with cell proliferation and survival.
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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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