Overview

  • Product name
  • Description
    Rabbit polyclonal to EDA2R
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length EDA2R (Human)

  • Positive control
    • HEK 293 (Human embryonic kidney cell line) whole cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab37171 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
IHC-P Use a concentration of 10 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).

Target

  • Function
    Receptor for EDA isoform A2, but not for EDA isoform A1. Mediates the activation of the NF-kappa-B and JNK pathways. Activation seems to be mediated by binding to TRAF3 and TRAF6.
  • Sequence similarities
    Contains 3 TNFR-Cys repeats.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 9430060M22Rik antibody
    • Ectodysplasin A2 isoform receptor antibody
    • Ectodysplasin A2 receptor antibody
    • EDA A2R antibody
    • EDA-A2 receptor antibody
    • EDA2R antibody
    • EDAA2R antibody
    • MGC124099 antibody
    • OTTMUSP00000019574 antibody
    • OTTMUSP00000019575 antibody
    • RP23-188N15.1 antibody
    • TNFRSF27 antibody
    • TNR27_HUMAN antibody
    • Tumor necrosis factor receptor superfamily member 27 antibody
    • Tumor necrosis factor receptor superfamily member XEDAR antibody
    • X linked ectodysplasin A2 receptor antibody
    • X linked ectodysplasin receptor antibody
    • X-linked ectodysplasin-A2 receptor antibody
    • XEDAR antibody
    see all

Images

  • Lane 1 : Anti-EDA2R antibody (ab37171) at 0.5 µg/ml
    Lane 2 : Anti-EDA2R antibody (ab37171) at 1 µg/ml
    Lane 3 : Anti-EDA2R antibody (ab37171) at 2 µg/ml

    All lanes : Human 293 cell lysate

    Lysates/proteins at 15 µg per lane.

    Predicted band size: 33 kDa
    Observed band size: 33 kDa

  • ab37171 at 10 µg/ml staining EDA2R in human skin tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections).

References

ab37171 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293 (human embryonic kidney))
Total protein in input
40 µg
Immuno-precipitation step
Protein A/G
Specification
HEK 293 (human embryonic kidney)

Abcam user community

Verified customer

Submitted Sep 30 2017

Answer

Thank you for taking the time to complete our questionnaire and send these images. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
The details you have kindly provided will enable us to investigate this case for you and also gives us vital information for our monitoring of product quality
I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. I would like to reassure you that this antibody is tested and covered by our six month guarantee for WB and human samples. Regrettably in this case the product was purchased in 2011 (longer than six month ago) and is for this reason it is not covered by this guarantee.
I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some details of the procedure.
1. Could you confirm which secondary antibody exactly was used for this experiment? I have never heard of a secondary antibody raised in human. It would be helpful if you could forward the catalogue number and company name where it was purchased. I would be pleased to compare the secondary for suitability.
2. We would recommend it might be beneficial to integrate a loading control within the experiment if not already performed. A loading control will ensure that the sample is of good quality. Furthermore it will assess the transfer of the sample to the membrane.
3. Additionally we would suggest performing a no primary control if not already done. This will show if background that is detected might be due to non-specific binding of the secondary antibody.
4. Sometimes the change of blocking solution can improve the result of a WB. We would suggest it might be beneficial trying to block with 3-5% BSA instead of milk. An example for how the blocking can affect the result is shown for our GAPDH antibody ab9385 here: www.abcam.com/ab9385 .
We are happy to offer this technical support. I hope this information may be helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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Question

Hello,
Here is a complain from one of the customer .
This antibody has been replaced earlier and that is replaced antibody supplied before a year ago .
1) Abcam catalogue no:
Ab37171

2) a) Abcam order number (not required):
958152 ,oct 3rd week,2011

b) Abcam product lot number:
GR53870-2

3) Description of the problem (high background, wrong band size, more bands, no band etc):
High background,more bands,no bands

4) Antibody storage conditions (temperature/reconstitution etc):
4oC

5) a)Sample (Species):
Human

b) Type of sample (Cell extract/Nuclear extract/Purified protein/Recombinant protein etc).:

Cell extract

6) a) Sample preparation:
Lysis buffer
Ripa buffer
Protease inhibitors
Sigma protease inhibitor cocktail
Phosphatase inhibitors
Sodium floride,sodium orthovernadate
Reducing agent
DTT
Boiling for ≥5 min?
5,10min
b) Amount of protein loaded:
Protein loaded ug/lane or cells/lane
60,70,80,100,200
7) a) Electrophoresis/Gel conditions:
Reducing or Non Reducing gel
Reducing gel
Percentage of gel
10%
Volts applied
120
Time applied
1.5-2hrs
b) Transfer or blocking conditions:
Type of membrane
Pvdf
Protein transfer verified
Yes
Blocking agent and concentration
Non fat dry milk,3-5%
Blocking time
1hr
Blocking temperature
RT
8) Primary antibody (If more than one was used, describe in “additional notes”):
Concentration or dilution
1000 times
Diluent buffer
Blocking in TBST
Incubation time
Overnight,3hrs
Incubation temperature
ON 40C,3hrs RT
Washing: Buffer Used
TBST
Number of washes
3
9) Secondary antibody:
Species
Human
Reacts against
Rabbit
Concentration or dilution
10000-15000 times
Diluent buffer
Blocking in TBST
Incubation time
1hr
Incubation temperature:
RT
Fluorochrome or enzyme conjugate
Enzyme conjugated
Washing: Buffer Used
TBST
Number of washes
3
10) Detection method (ECL, ECL plus, etc.):
ECL
11) Positive and negative controls used:
Positive control
293T cells
Negative control
12) a) How many times you have run this staining?
5-7 times
b) Do you obtain the same results every time?
No it varies,from totally dark back ground to ,many non specific bands(without any specific band) or a totally clear blot.
C) What steps have you altered to try and optimize the use of this antibody?
Tried with different protein concentration,different dilution of primary and secondary antibody,different cell lines.secondary antibodies from two different companies.
Regards

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.
I would like to reassure you that ab37171 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement. We are also pleased to provide technical support to help to improve the result.
Reviewing this case, I would appreciate if you can send some of your different WB results. Could you confirm, what the differences were performing this WB experiments?
Thank you for your cooperation. I look forward hearing from you with the further requested details.

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Answer

Thank you very much for your interest in ab37171. To our knowledge, ab37171 has not been tested in mouse. Therefore, I can offer a discount off a future purchase if you buy ab37171 now, test it in mouse 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 ab37171 in mouse. I will then send a discount code. This code must be issued before purchasing ab37171 so please wait for my reply before ordering. 2. Purchase ab37171 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse. 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 more about our Abreview system, 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 any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab37171 turns out to be unsuitable for mouse, 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.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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