Overview

  • Product name
    Anti-SET7 antibody [s4E5]
    See all SET7 primary antibodies
  • Description
    Mouse monoclonal [s4E5] to SET7
  • Host species
    Mouse
  • Specificity
    This antibody has been tested against recombinant SET7 but has not been tested against the endogenous protein yet.
  • Tested applications
    Suitable for: ELISA, WB, ChIP, IHC-Glut, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein corresponding to Human SET7.

  • Positive control
    • Recombinant SET7

Properties

Applications

Our Abpromise guarantee covers the use of ab14820 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
ELISA Use at an assay dependent concentration.
WB 1/250 - 1/2000. Predicted molecular weight: 50 kDa.
ChIP Use at an assay dependent concentration. PubMed: 21859860
IHC-Glut Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Histone methyltransferase that specifically monomethylates 'Lys-4' of histone H3. H3 'Lys-4' methylation represents a specific tag for epigenetic transcriptional activation. Plays a central role in the transcriptional activation of genes such as collagenase or insulin. Recruited by IPF1/PDX-1 to the insulin promoter, leading to activate transcription. Has also methyltransferase activity toward non-histone proteins such as p53/TP53, TAF10, and possibly TAF7 by recognizing and binding the [KR]-[STA]-K in substrate proteins. Monomethylates 'Lys-189' of TAF10, leading to increase the affinity of TAF10 for RNA polymerase II. Monomethylates 'Lys-372' of p53/TP53, stabilizing p53/TP53 and increasing p53/TP53-mediated transcriptional activation. Also able to demethylated 'Lys-372' of p53/TP53 in vitro.
  • Tissue specificity
    Widely expressed. Expressed in pancreatic islets.
  • Sequence similarities
    Belongs to the histone-lysine methyltransferase family. SET7 subfamily.
    Contains 3 MORN repeats.
    Contains 1 SET domain.
  • Domain
    The SET domain is necessary but not sufficient for histone methyltransferase activity.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ21193 antibody
    • H3 K4 HMTase antibody
    • H3-K4-HMTase SETD7 antibody
    • H4 lysine 4 specific antibody
    • Histone H3 K4 methyltransferase antibody
    • Histone H3 lysine 4 specific methyltransferase antibody
    • Histone H3-K4 methyltransferase SETD7 antibody
    • Histone H4 K4 methyltransferase antibody
    • Histone lysine N methyltransferase antibody
    • Histone lysine N methyltransferase H3 lysine 4 specific SET7 antibody
    • Histone-lysine N-methyltransferase SETD7 antibody
    • KIAA1717 antibody
    • KMT7 antibody
    • Lysine methyltransferase antibody
    • Lysine N-methyltransferase 7 antibody
    • OTTHUMP00000164543 antibody
    • OTTHUMP00000220049 antibody
    • SET 7 antibody
    • SET 7/9 antibody
    • SET 9 antibody
    • SET D7 antibody
    • SET domain containing (lysine methyltransferase) 7 antibody
    • SET domain containing protein 7 antibody
    • SET domain containing protein 8 antibody
    • SET domain-containing protein 7 antibody
    • SET7 antibody
    • SET7/9 antibody
    • SET9 antibody
    • Setd7 antibody
    • SETD7_HUMAN antibody
    see all

Images

  • Western blot analysis of recombinant SET7 using ab14820 at 1/1000. Proteins were visualised using a goat anti-mouse secondary antibody conjugated to HRP and a DAB detection system. Western blot analysis of recombinant SET7 using ab14820 at 1/1000. Proteins were visualised using a goat anti-mouse secondary antibody conjugated to HRP and a DAB detection system.
  • Immunoflouresence was carried out using mouse cells in culture. Conditions: 4% PFA fixation and dilution with 0.5% triton, 10mg/ml BSA in PBS. Dilution of ab14820 used: 1/50. Green = ab14820 detected with secondary antibody (Alexa 488). Blue = DAPI stain.
  • ab14820 at 1/25 dilution staining SET7 / KMT7 / SETD7 / SET7/9 in mouse pancrease tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed prior to blocking in 5% serum for 1 hour at 20°C and then incubated with ab14820 for 18 hours at 4°C. A Cy2 conjugated donkey polyclonal to mouse Ig, diluted 1/20, was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab14820 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14820, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Xu X  et al. A signature motif in LIM proteins mediates binding to checkpoint proteins and increases tumour radiosensitivity. Nat Commun 8:14059 (2017). ChIP ; Mouse . Read more (PubMed: 28094252) »
  • Zhang Y  et al. The transcription factor GATA1 and the histone methyltransferase SET7 interact to promote VEGF-mediated angiogenesis and tumor growth and predict clinical outcome of breast cancer. Oncotarget 7:9859-75 (2016). Read more (PubMed: 26848522) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Answer

Thank you for your continuous interest in our product.

ab14820 has already been tested and characterized for ChIP application so our classical testing Abtrial (for new untested application or species) does not qualify for the testing discount.

However, since we do not have currently a ChIP image of ab14820 we can invite you for an image Abtrial.

By participating in our Image AbTrial program you can now use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit an image of your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

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Question
Answer

Thank you for your kind reminder for ab13731 and for your patience.

The re-testing has been finished. This antibody (ab13731: Anti-SETD7 antibody - ChIP Grade) was tested on both MCF-7 and HeLa cell lysate. Please see attached results. A single main band of the predicted was seen in MCF-7 cells and several bands in HeLa lysate. Our standard WB protocol was used to generate the results.

The variability of banding patterns could be due to differential expression between cell types and lysate preparations. With any given antibody, it is not uncommon that different WB banding patterns are seen between different cell types or different lysate preparations of the same cell type.

At this stage, it can be hard to know the significance of this but it could be due to differential expression of isoforms, post-translational modifications, cleavages in different cell types or different phases of the cell cycle. It could also be due to “artefacts” of the cell lysis.

I am not sure which situation may apply to the results (differences in banding patterns between MCF-7 and HeLa cell lysate) as well as that of your customer. You may want to compare WB results between different preparations of the cell lysate.

Please let me know if you have any additional questions.

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Answer

Thank you for your prompt answer and for answering to my further questions.

This is to confirm that we are going to retest the antibody in HeLa cell lysate and I should have the results by the end of this week or early next week.

I will update you as soon as possible. Thank you for your patience in advance!

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Question

Thanks for the credit code for antibody ab14820 (SETD7). I applied this to purchase another Abcam antibody against SETD7 (ab13731). However, I am really disappointed as it is no better than ab14820. SETD7 is 43 kDa and as you can see in the positive control HEK293 there is no reactivity at this molecular weight, rather a major blot is seen at about 140kD.

Do you have any suggestions for improving the result. I have attached the western blot and used the following conditions:

Sample preparation:

Type of sample : whole cell lysate from 16HBE14o- and Caco2; nuclear protein from HEK293

Lysis buffer : RIPA buffer

Protease inhibitors: Roche ecomplete proteinase inhibitor cocktail and PMSF

Phosphatase inhibitors

Reducing agent : ß-mercaptoethanol and SDS

Boiling for ≥5 min? yes

Protein loaded ug/lane or cells/lane: about 30 ug lysate each lane

Positive control : I tried IRF2 western blotting at the same, which is a rabbit polyclonal antibody.

Negative control I have one lane left only running the dye.



Percentage of gel 8%

Type of membrane: PVDF membrane

Protein transfer verified:Yes, Ponceau Red

Blocking agent and concentration: 5% nonfat milk

Blocking time 1hr

Blocking temperature Room temp



Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 1:2000 (1st time); 1:500 (2nd time)

Diluent buffer 5% nonfat milk

Incubation time overnight

Incubation temperature:4 degree in cold room


Washing after primary and secondary antibodies:

Buffer 1XTBST buffer

Three times, 10-15 min each


How many times have you run this staining?

Twice

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Answer

Thank you for your enquiry regardingab13731 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

The other antibody (ab14820) you have tried was only tested on recombinant protein but not on endogeously expressed system. However, ab13731 has been characterized for Western blot application on HeLa cell lysate and a clear and distinctive band was obtained at the expected size (43 kDa).

The protocol details you have kindly sent to me looks fine. I would like to make the following comments/suggestions:

1) Secondary antibody and detection:

What is the host species of the secondary antibody and what type of immunoglobulin was it raised against?





Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not? If you have not done yet, I would advise you to check it.



2) Blocking:

Have you tried other blocking agent like 5 % BSA in TBS-T rather than milk which for certain target proteins works better?



3) Lysis buffer:

It may be worth peparing some new cell lysate with freshly prepared lysis buffer.

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

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Question
Answer

Vielen Dank für Ihre Antwort.

Es tut mir leid, dass die Vorschläge das Problem nicht lösen konnten. Ich muss daraus schliessen, dass das Röhrchen welches Sie bekommen haben beschädigt ist. Es tut uns sehr leid. Wir senden Ihnen natürlich gerne einen Ersatz. Sie können wählen, ob Sie als Ersatz ein neues Röhrchen des ab14820 hätten, ein Röhrchen eines alternativen Antikörpers oder eine Gutschrift/Rückerstattung. Können Sie uns bitte hierzu auch die originale Bestellnummer des ab14820 geben? Falls Sie die Nummer nicht haben, können Sie uns das Datum der Bestellung sowie den Namen der Person die den ab14820 bestellt hat geben? Vielen Dank.

Leider haben wir zur Zeit jedoch keinen Setd7 Antikörper in unserem Katalog, der unseres Wissens nachauf Frosch oder Fischgewebe getestet wurde. Der ab124708 ist der einzige Antikörper gegen Setd7, der schon in IP und WB getestet wurde. Ich habe nun das Labor angefragt, ob die Immunogensequenz gewisse Homologie mit der Proteinsequenz von Setd7 von Fisch und Frosch hat. Ich werde mich melden sobald ich eine Antwort habe.

Unabhängig von der Ersatzsendung, können wir Ihnen dann auch ein Testangebot machen, falls Sie einen anderen Setd7 Antikörper von uns auf Fisch und Frosch testen wollen. Details eines solchen Angebots sind hier zu finden: https://www.abcam.com/collaborationdiscount

Ich freue mich wieder von Ihnen zu hören.

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Question
Answer

Vielen Dank für Ihre Email und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Um die Ursache des Problems herauszufinden undzu lösen,möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen. Ich wäre Ihnen auch dankbar, wenn Sie gewisse Punkte genauer erklären könnten.

1.) Können Sie bitte bestätigen, dass das rekombinantefull length Protein benutzt wurde?

2.) Wissen Sie, ob in den Western blot lysaten genug rekombinantes oder endogenes Protein vorhanden ist, sodass es detektiert werden kann?

3.) Ich kann Ihnen auch empfehlen, BSA als Blockierungsmittel für den Western blot auszuprobieren. In der Tat, kann ein anderer Blockierungspuffer das Resultat signifikant verändern. Als Beispiel dafür, hat es ein Bild auf dem Datenblatt von ab9385 https://www.abcam.com/index.html?datasheet=9385 (or use the following: https://www.abcam.com/index.html?datasheet=9385).

4.) Für die Immunhistochemie, kann ich Ihnen emfpehlen die gefrorenen Schnitte nur 10 Minuten mit 4%PFA zu fixieren. Dies ist genug für normale Gefrierschnitte. In der Tat kann eine zu lange Fixierung das Gewebe so verändern, dass es nicht mehr gefärbt werden kann. Wenn 10 Minuten fixiert wird, brauchen Sie auch keine Demaskierung. Sie können danngleich mit der Permeabilisierung fortfahren. Bitte lassen Sie mich wissen, falls ich Ihr Protokoll missverstanden habe.

Wie schon gesagt, falls wir das Problem nicht lösen können, werden wir Ihnen diesen Antikörper ersetzen.

Ich freue mich wiedervon Ihnen zu hören.

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Answer

Thank you for contacting us.

Your credit note ID is 20804.

I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

Vielen Dank für Ihren Anruf.
Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.
Wei am Telefon diskutiert, sende ich Ihnen hier unsere Fragebogen. Es tut mir leid, dass ich gestern nicht mehr dazu gekommen bin. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.
Ich freue mich, bald wieder von Ihnen zu hören.

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Question

Thank you for the quick reply.The email and attachment includes all the information I could provide. Can you please give me some suggestions about how can I fix my experiment? Any suggestions are welcomed. I look forward to hearing from you soon and helping me solve the problem.


Could you please confirm if you purchased this product directly from Abcam or via a distributor?



Yes. I purchased directly form Abcam.



It would be very useful for me to know the Abcam Order Number (or your PON) and the date of the purchase.

Purchase order # 53949316

Receipt Date: 04-20-2012





1) Abcam product code ab14820



2) Abcam order reference number or product batch number Lot. GR18771-8



3) Description of the problem

The blots are not shown as the datasheet. Please see the photos I sent.



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…) whole cell lysate from 16HBE14o- and Caco2

Lysis buffer : RIPA buffer

Protease inhibitors: Roche ecomplete proteinase inhibitor cocktail and PMSF

Phosphatase inhibitors

Reducing agent : ß-mercaptoethanol and SDS

Boiling for ≥5 min? yes

Protein loaded ug/lane or cells/lane: about 30-40 ug lysate each lane

Positive control : Both cell lysates have been tested with anti-SETD7 from other company giving the correct size product( 50kDa). On the same gel I have run the same samples for anti-NFYA (Abcam), GR(Santa cruz) at the same time. They are all right.

Negative control I have one lane left only running the dye.



5) Percentage of gel 8%

Type of membrane: PVDF membrane

Protein transfer verified:Yes, Ponceau Red

Blocking agent and concentration: 5% nonfat milk

Blocking time 1hr

Blocking temperature Room temp



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 1:2000

Diluent buffer 5% nonfat milk

Incubation time overnight

Incubation temperature:4 degree in cold room



7) Secondary antibody:

Species: Polyclonal Goat Anti-Mouse Immunoglobulins/HRP,Code No./ Code/ Code-Nr. P 0447

Reacts against: mouse

Concentration or dilution 1:5000

Diluent buffer 5% nonfat milk

Incubation time 1hr

Incubation temperature: Room temp

Fluorochrome or enzyme conjugate:



8) Washing after primary and secondary antibodies:

Buffer 1XTBST buffer

Number of washes three times, 10-15 min each



9) Detection method



10) How many times have you run this staining?

Once only.

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Answer

Thank you very much for getting back to me and for passing some useful details. I have looked through the protocol and it seems to be fine.

I have discussed this complaint with my colleagues in the Lab and this antibody has only been tested on Recombinant SET7 system but we do not have any data on endogenously expressed system cell or tissue lysate.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

My suggestion would be to try increasing the final working concentration of the primary antibody i.e. 1/1000 or 1/500 (overnight incubation) to see if the signal is getting stronger or not. If this does not improve the detection, I can certainly arrange the credit note for you or send you a replacement.

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

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Answer

Thank you for your enquiry regardingab14820 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

Could you please confirm if you purchased this product directly from Abcam or via a distributor? It would be very useful for me to know the Abcam Order Number (or your PON) and the date of the purchase.

Would you be so kind to provide some further details of the protocol used and complete the following form (attached as a word document).

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

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1-10 of 15 Abreviews or Q&A

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