Overview

  • Product name

    Anti-ZNF420 antibody
  • Description

    Rabbit polyclonal to ZNF420
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Horse, Cat
  • Immunogen

    Synthetic peptide corresponding to a region within N terminal amino acids 36-85 (LENYSNLVSL DLPSRCASKD LSPEKNTYET ELSQWEMSDR LENCDLEESN) of Human ZNF420 (NP_653290).

  • Positive control

    • 293T cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab84618 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 80 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Relevance

    ZNF420 contains 19 C2H2-type zinc fingers and 1 KRAB domain. It may be involved in transcriptional regulation. It is a negative regulator of p53-mediated apoptosis.
  • Cellular localization

    Nuclear
  • Database links

  • Alternative names

    • APAK antibody
    • ATM and p53 associated KZNF protein antibody
    • FLJ32191 antibody
    • Zinc finger protein 420 antibody
    • ZNF 420 antibody
    see all

Images

  • Anti-ZNF420 antibody (ab84618) at 1 µg/ml (in 5% skim milk / PBS buffer) + 293T cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 80 kDa
    Observed band size: 80 kDa



    Gel concentration: 12%

References

This product has been referenced in:

  • Yu B  et al. Diagnostic potential of serum exosomal colorectal neoplasia differentially expressed long non-coding RNA (CRNDE-p) and microRNA-217 expression in colorectal carcinoma. Oncotarget 8:83745-83753 (2017). Read more (PubMed: 29137379) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for confirming those details.

I have arranged for a new vial of ab84618 to be sent to you. This is on the order number xxxxx(Purchase order number FOCR xxxxx). Unfortunately this is not currently in stock and it may take up to two weeks to reach you. I am sorry for the inconvenience this may cause you.

If you would be happy to provide me with the results you obtained with the previous lot that would be very helpful in investigating the problem further. I have attached a questionnaire to this email for you to fill out if you wouldn't mind. This should only take 5 minutes and if you could include some of the images obtained that would bevery useful.

I look forward to receiving your reply.

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Answer

Thank you for getting back to me with those details.

I have located an order of the 23rd of April delivered to the following address:
xxxxxxxx

Are these details correct?

I can reassure you that if you find the same problems with the replacement antibody I would try my best to try and resolve the problem. If I was unable to provide technical advice as to what may be contributing to the problems you would be entitled to a free replacement antibody of your choice or a refund.

I look forward to receiving your reply.

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Question

1) Abcam product code ab84618
2) Abcam order reference number or product batch number: GR82869-1
3) Description of the problem: I recently purchased a ZNF420 antibody (ab84618) from Abcam. This antibody worked well for the first blot that I used it for and since then has stopped working appropriately even though I have stored and used it as recommended.
4) Sample preparation:Type of sample: Whole cell extract
Lysis buffer: UTB lysis buffer: 9M urea, 75mM Tris-HCl pH 7.5 and 0.15M -mercaptoethanol
Boiling for ≥5 min? yes
Protein loaded ug/lane or cells/lane between 30-50 ug/lane loaded. Cells testet include primary cell lines (WI38), and colorectal cancer cell lines (RKO and HCT116)
Positive control: It has previously been published that HCT116 cells have good expression levels of ZNF420, therefore the lysates from these cells was used as a positive control. No ZNF420 expression, however, was observed in these lysates using this abcam antibody.
5) Percentage of gel Gradient 4-20% gel usedType of membrane: Nitrocellulose 0.45 um
Protein transfer verified: transfer was successful as seen by ponceau staining
Blocking agent and concentration; Lycor buffer in PBS (1:1) was used as blocking agent for 1 hour at room temperature
6) Was a fresh membrane used or had it been probed and stripped with a different antibody? Fresh membranes were used.
7) Primary antibody (If more than one was used, describe in “additional notes”) :Concentration or dilution 1:500
Diluent buffer: Lycor buffer in PBS-T (0.1%) (1:1)
Incubation time: Overnight
Incubation temperature: 4oC
8) Secondary antibody:
Species: Rabbit
Reacts against: Human
Concentration or dilution 1:10000
Diluent buffer: Lycor buffer in PBS-T (0.1%) (1:1)
Incubation time 1-2 hours
Incubation temperature: Room temperature
Fluorochrome or enzyme conjugate:680
9) Washing after primary and secondary antibodies:Buffer PBS-T (0.1%)
Number of washes 3 after primary antibody (5 minutes each) and 3 after secondary antibody (last wash in PBS to reduce background when scanning)
10)Detection method: Odyssey system
1) How many times have you run this staining? 4
Do you obtain the same results every time? Antibody worked well for the first time the staining was tried. The second time the antibody was already working less well. It has not worked ever since, even in those cells that are know to have good expression levels of the protein
What steps have you altered to try and optimize the use of this antibody? I have increased the amount of protein loaded per well. I have used fresh primary and secondary antibodies, I have used pre-cast gels and I have used cell lines with known good expression and this has not seemed to help with the problem.

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Answer

Thank you for taking time to complete our questionnaire. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been more successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. May I ask, when did you purchase this antibody? Do you have the order number available (or the delivery address)? Would it be possibleto share an imageof the blotwhich did work with the antibody?

I apologise for the inconvenience this problem has caused you. I would be pleased to offer you a free of charge replacement of this antibody (or a different one of your choice from our catalogue). Alternatively I can arrange for a credit note or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with the anti-ZNF420 antibody (ab84618). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I will be looking into if we have any alternative lots of this antibody available, as well as if we have had any similar problems reported. In order to investigate the problem further, would you mind providing me with a few more details of you experiments? I have attached a questionnaire to this email. If you wouldn't mind filling it in and including any images you have of the blots performed that would be very helpful in understanding the problem more fully.

If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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