Overview

  • Product name
    Anti-EHMT1/GLP antibody [B0422]
    See all EHMT1/GLP primary antibodies
  • Description
    Mouse monoclonal [B0422] to EHMT1/GLP
  • Host species
    Mouse
  • Tested applications
    Suitable for: ChIP, ICC/IF, WB, ELISA, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to Mouse EHMT1/GLP aa 134-234.
    Database link: Q5DW34

  • Positive control
    • WT mouse ES cell

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purification notes
    ab41969 was purified by Ammonium sulfate fractionation
  • Clonality
    Monoclonal
  • Clone number
    B0422
  • Myeloma
    NS1
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab41969 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration. PubMed: 18066052
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 0.5 µg/ml. Predicted molecular weight: 134 kDa.
ELISA Use a concentration of 0.1 µg/ml.
IP Use at an assay dependent dilution.

Target

  • Function
    Histone methyltransferase that specifically mono- and dimethylates 'Lys-9' of histone H3 (H3K9me1 and H3K9me2, respectively) in euchromatin. H3K9me represents a specific tag for epigenetic transcriptional repression by recruiting HP1 proteins to methylated histones. Also weakly methylates 'Lys-27' of histone H3 (H3K27me). Also required for DNA methylation, the histone methyltransferase activity is not required for DNA methylation, suggesting that these 2 activities function independently. Probably targeted to histone H3 by different DNA-binding proteins like E2F6, MGA, MAX and/or DP1. During G0 phase, it probably contributes to silencing of MYC- and E2F-responsive genes, suggesting a role in G0/G1 transition in cell cycle. In addition to the histone methyltransferase activity, also methylates non-histone proteins: mediates dimethylation of 'Lys-373' of p53/TP53.
  • Tissue specificity
    Widely expressed.
  • Involvement in disease
    Defects in EHMT1 are the cause of chromosome 9q subtelomeric deletion syndrome (9q- syndrome) [MIM:610253]. Common features seen in these patients are severe mental retardation, hypotonia, brachy(micro)cephaly, epileptic seizures, flat face with hypertelorism, synophrys, anteverted nares, cupid bow or tented upper lip, everted lower lip, prognathism, macroglossia, conotruncal heart defects, and behavioral problems.
  • Sequence similarities
    Belongs to the histone-lysine methyltransferase family.
    Contains 8 ANK repeats.
    Contains 1 pre-SET domain.
    Contains 1 SET domain.
  • Domain
    The ANK repeats recognize and bind RELA subunit of NF-kappa-B, when RELA is monomethylated at 'Lys-310' (By similarity). They also specifically recognize and bind H3K9me1 and H3K9me2.
    The SET domain mediates interaction with WIZ.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Nucleus. Chromosome. Associates with euchromatic regions.
  • Information by UniProt
  • Database links
  • Alternative names
    • bA188C12.1 antibody
    • DKFZp667M072 antibody
    • EHMT 1 antibody
    • EHMT1 antibody
    • EHMT1_HUMAN antibody
    • Eu-HMTase1 antibody
    • Euchromatic histone lysine N methyltransferase 1 antibody
    • Euchromatic histone methyltransferase 1 antibody
    • Euchromatic histone-lysine N-methyltransferase 1 antibody
    • EUHMTASE1 antibody
    • FLJ12879 antibody
    • FP13812 antibody
    • G9a like protein 1 antibody
    • G9a like protein antibody
    • G9a-like protein 1 antibody
    • GLP 1 antibody
    • GLP antibody
    • GLP1 antibody
    • H3 K9 HMTase 5 antibody
    • H3-K9-HMTase 5 antibody
    • Histone H3 K9 methyltransferase 5 antibody
    • Histone H3-K9 methyltransferase 5 antibody
    • Histone lysine N methyltransferase H3 lysine 9 specific 5 antibody
    • Histone-lysine N-methyltransferase EHMT1 antibody
    • KIAA1876 antibody
    • Lysine N methyltransferase 1D antibody
    • Lysine N-methyltransferase 1D antibody
    • RP11 188C12.1 antibody
    see all

Images

  • Ab41969 at 5 µg/ml staining WT mouse ES cell (Positive)
  • Ab41969 at 5µg/ml staining GLP KO mouse ES cell (Negative)

References

This product has been referenced in:
  • Li H  et al. Epigenetic control of IL-23 expression in keratinocytes is important for chronic skin inflammation. Nat Commun 9:1420 (2018). Read more (PubMed: 29650963) »
  • Yang Y  et al. Euchromatic histone lysine methyltransferase 1 regulates cancer development in human gastric cancer by regulating E-cadherin. Oncol Lett 15:9480-9486 (2018). Read more (PubMed: 29844834) »
See all 14 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Dermal Microvascular Endothelial Cells)
Permeabilization
Yes - 0.1% Triton-X100
Specification
Human Dermal Microvascular Endothelial Cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 13 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (B cells)
Negative control
IgG
Specification
B cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
Gene Promoter

Abcam user community

Verified customer

Submitted Apr 27 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - nuclear (Dermal Endothelial Cells (Primary))
Total protein in input
300 µg
Immuno-precipitation step
Protein A/G
Specification
Dermal Endothelial Cells (Primary)

Abcam user community

Verified customer

Submitted Apr 24 2017

Application
Western blot
Sample
Human Cell lysate - nuclear (Dermal Endothelial cells (Primary))
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
Dermal Endothelial cells (Primary)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 21 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (C2C12 myoblast cells)
Permeabilization
No
Specification
C2C12 myoblast cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 07 2013

Answer

Thank you so much for your reply.
Below are the testing discount codes for both GLP antibodies. At the bottom of the email is the information about the free of charge replacement for ab41969. A PO number is not needed on my end for sending you the codes and the replacement.
DISCOUNT CODE: xxx for ab63161
DISCOUNT CODE: xxx for ab84751
Expiration date: xxx
I am very pleased to hear you would like to accept our offer and test ab63161 and ab84751 in mouse and/or WB. Each code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for each antibody for mouse and/or WB, and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.
*** FREE OF CHARGE REPLACEMENT Information ***
Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab63161. The estimated delivery date is February 28th, 2012.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Answer

Thank you for your reply.
Yes, I think you are understanding this correctly.
I will be able to replace ab41969 free of charge with one vial of either ab63161 or ab84751. I am not able to send you both. Unfortunately, we do not offer sample sizes of our products.
For whichever antibody you select (ab63161 or ab84751) I will also send you a testing discount code since neither of them have been tried in mouse and/or WB. Neither of them would be covered under our guarantee - but that's why we give out the testing code.
Independent of whether the selected antibody works or not, you will receive a free antibody on your next order - once you have submitted your data in form of an Abreview (from the online datasheet of the antibody or your Abcam account).
With the code you could purchase the second GLP antibody, but another testing discount could not be given out in that case.
I can, however, give you testing discount code for trying the second GLP antibody which you would need to purchase at full price.
The price of each antibody is:
ab63161 100µg £227.00 + £16.00 shipping (available on Feb 28 if ordered today)
ab84751 100µl £227.00 + £16.00 shipping (currently backordered for 1-2 weeks)
TESTING DISCOUNT INFORMATION:
For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
I would need the following information from you:
1) Please let me know which antibody I shall send you free of charge (+ testing discount code): ab63161 or ab84751?
2) Would you like me to send you a testing discount code for the second GLP antibody as well?
I look forward to hear back from you and resolve this case for you.

Read More

Question

Thanks for the rapid response. First I didn't find any problem with the ab1220 only ab41969. I mention it only because I've used it to run gels on all of the same samples (more samples than necessary) as a control and to show (indirectly) GLP/G9a-methylation present in the brain during the embryonic stages I was examining. So in effect, I've used a lot of this antibody as a result of ab41969 not working.

In response to your questions:

1. I used RIPA buffer (following abcams protocol)
RIPA buffer (Radio Immuno Precipitation Assay buffer)

150 mM sodium chloride

1.0% NP-40 or Triton X-100

0.5% sodium deoxycholate

0.1% SDS (sodium dodecyl sulphate)

50 mM Tris, pH 8.0


2. I digested whole brains for E12.5-P1 at 5mg/1ml of lysises buffer; after diluting with the SDS lysis buffer I was loading 10uL of sample (which was about 30 ug of protein).

3. I've tried numerous concentrations of my antibody (from 1:250-1:2000), which were close in range to the suggest dilution. And I've tried 2 and 4hrs incubation on primary with 1 hour on secondary.

4. Yes, as mentioned I was running a control with all the same materials/protein/gel/samples with ab1220 to be certain the stages were showing high methylation, and subsequently would expect high GLP concentrations. Subsequently ab1220 was working very efficiently. And Both antibodies are monoclonal and the same secondary was being used.

5. TBS- with tween and with 5% milk

Instead of a refund, it would be ideal to get another batch. In addition and as you mention, it would also be very useful to get the testing codes to try the untested antibodies. I'm happy to submit the results. The goal is to find a working antibody and this seems like the most efficient and timely way forward.

Read More
Answer

Thank you for your reply and the additional information which will be very helpful to us to evaluate the antibody.
I have checked, but unfortunately we do not have another new lot for ab41969 in stock right now. Instead, I can send you a replacement with either ab63161 (not tested in mouse) or ab84751 (not tested in mouse and WB) including a testing discount code.
Please let me know if which antibody you would prefer.

I look forward to hear back from you and resolve this case for you.

Read More

Answer

Thank you for contacting us.
I am very sorry to hear that the antibody ab41969 is giving you problems.
Could you please let me know the following details for this antibody:
1) How were your samples prepared (e.g. lysis buffer)?
2) How much protein was loaded?
3) What antibody dilutions and incubation time were tired?
4) I assume that the secondary antibody was working well otherwise?
5) What was used for blocking?
6) What is your order or PO number?
I'd be happy to replace ab41969 with either another lot or a different antibody. Unfortunately, none of the other GLP antibodies have been tested in mouse so far (ab63161) and ab84751 has also not yet been tested in WB. Therefore, we would not be able to guarantee them for this species/application. But if you would like to try, I can send you in addition a testing discount code (see below for details).
For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
Alternatively, a credit or refund can be arranged.
Please let me know how you would like to proceed with ab41969.
As for ab1220: What problems did you encounter with this antibody? Please let me know in detail as well.
I look forward to hear back from you.

Read More

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