Overview

  • Product name

  • Description

    Goat polyclonal to Npas4
  • Host species

    Goat
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Dog, Pig, Zebrafish
  • Immunogen

    Synthetic peptide:

    CRFNTSKSLRRQS

    , corresponding to internal sequence amino acids 149-161 of Human Npas4 (NP_849195.2).

  • Positive control

    • Human brain (frontal cortex) lysate

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: 99% Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab109984 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 0.3 - 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 87 kDa).

Target

Images

  • All lanes : Anti-Npas4 antibody (ab109984) at 1 µg/ml

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 87 kDa

  • Anti-Npas4 antibody (ab109984) at 1 µg/ml + Human frontal cortex lysate (in RIPA buffer) at 35 µg

    Developed using the ECL technique.

    Predicted band size: 87 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 hour

References

This product has been referenced in:

  • Zhang Z  et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). WB ; Mouse . Read more (PubMed: 30297870) »
  • Liu L  et al. Hippocampal Mechanisms Underlying Impairment in Spatial Learning Long After Establishment of Noise-Induced Hearing Loss in CBA Mice. Front Syst Neurosci 12:35 (2018). Read more (PubMed: 30087600) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you again for your enquiry


I am sorry that you are having problems with this antibody. Unfortunately, it looks like we are not able to investigate this further, because it may be that the antibody may be working but is not able to detect the protein in an untested species.


Our policy is that we are happy to offer a refund, credit note or free of charge replacement when a product is not working in a successfully tested applications or species (and the product has been purchased in the last 180 days).That means as you are working with a non-tested speciesthat we cannotreplace the antibody.


To help you further with your case, I would like to make the following comments to the answers you kindly provided:


1. There are already prepared human tissue lysates available from different vendors, including us. If you are working with an untested species, it is recommended to run a positive control to asses how well the antibody is working.

2. It is not clear what conditions are used within the gel system. Most of our antibodies are tested and optimised to work under reducing conditions, and we usually state on our datasheet if the gel should be run under non-reducing conditions. I would therefore suggest to run a SDS-PAGE under reducing conditions. Otherwise it may be that the antibody may not be able to detect its target.

3. beta- Actin and GAPDH are internal controls on how well the proteins between 36 and 42 kDa have been transferred, but not a more heavier protein like Npas4. In addition, these two proteins are rather found outside the nucleus, so running a nuclear loading control like TATA binding protein TBP would be more advisable as well. Please check with Ponceau as these twoproteins do notindicate that the transfer is evenly above 42 kDa.


However, to solve this tricky situation, I can offer a discount off a future purchase if you buy ab109984 Anti-Npas4 antibody now, test it again in mouse and/orrat and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

If you are interested in this offer, please follow these steps:


1. Reply to this e-mail to let me know that you would like to proceed and test ab109984 in mouse and/or rat. I will then send a discount code. This code must be issued before purchasing ab109984 so please wait for my reply before ordering.

2. Purchase ab109984 either by phone, fax, or online (https://www.abcam.com or your distributor).

3. Test it in mouse and/or rat.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab109984 turns out to be unsuitable for mouse and/or rat, you will still receive the discount on your next purchase after your Abreview has been submitted.


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/collaborationdiscount.

Read More

Question

We have a quality issue of item ab109984. Please check attachment, and fix it for us.


1. Order details:
- Antibody storage conditions (temperature/reconstitution etc)
-20℃
2. Please describe the problem (high background, wrong band size, more bands, no band etc).
No band
3. On what mtaerial are you testing the antibody in WB?
* Species: rat & mouse
* What cells or tissue: hippocampi
* Cell extract or Nuclear extract:
* Purified protein or Recombinant protein:
4. The lysate
- How much protein was loaded: 60-80μg
- What lysis buffer was used: RAPA lysis buff
- What protease inhibitors were used:PMSF、Protease Inhibitor Cocktail、All-in-one (Protein phosphatase inhibitors mixture)
- What loading buffer was used: 1×SDS-PAGE loading buffer
- Did you heat the samples: temperature and time:100℃ 5min

5. Electrophoresis/Gel conditions/ Transfer conditions
- Reducing or non reducing gel:
- Gel percentage : 8%&10% separation,5%spacer gel
- Transfer conditions: 100 mA 3h (0℃) or 100 mA 2h (0℃) or 300 mA 1.5h (0℃) or 320 mA 1.5h (0℃)

6. Blocking conditions
- Buffer: TBS-T
- Blocking agent: milk, BSA, serum, what percentage: 5% milk or 5%BSA
- Incubation time: 1h or 1.5h
- Incubation temperature: RT
7. Primary Antibody
- Specification (in which species was it raised against):
* At what dilution(s) have you tested this antibody: 1:50&1:100&1:200&1:500
* What dilution buffer was used: 5% milk & 5%BSA
* Incubation time: over night(4℃) or 2h(RT)
* Incubation temperature: over night(4℃) or 2h(RT)
* What washing steps were done:10min×3times
8. Secondary Antibody
- Specification (in which species was it raised against)?
- At what dilution(s) have you tested this antibody:1:5000\1:7500\1:10000
- What dilution buffer was used: 5% milk
- Incubation time:1h\1.5h
- Wash steps: 7min×5times \10min×3times
- Do you know whether the problems you are experiencing come from the secondary?
9. Detection method
ECl, ECl+, other detection method:
10. Background bands
* Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):
* Is the blocking step sufficient?
* Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps)
* At what size are the bands migrating? Could they be degradation products of your target?
* Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient)
11. Did you apply positive and negative controls along with the samples? Please specify.
12. Optimization attempts
* How many times have you tried the Western? More than 3 times
* Do you obtain the same results every time e.g. are background bands always in the same place?
There was no band at all
* What steps have you altered?

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.


Please be aware that we haven't tested the species you are working with yet, they are only predicted to react due to their sequence homology. Unfortunately, this means that they are not covered by our guaranty.


Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab109984 Anti-Npas4 antibody. I would also appreciate if you can confirm some further details:


1. As these two species are not tested please run a positive control of human frontal cortex lysate to asses if the antibody is working.


2. Are you running the gel under reducing or native conditions?


3. Are you checking the transfer? How?


4. Please clarify "What dilution buffer was used: 5% milk & 5%BSA"
Are you using 5% milk & 5%BSA at the same time?


5. What kind of secondary antibody are you using?



Should the suggestions not improve the results, please do let me know.



I hope this information is helpful, and I thank you for your cooperation.

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