Overview

  • Product name

  • Description

    Rabbit polyclonal to EMR2
  • Host species

    Rabbit
  • Specificity

    ab75190 detects endogenous levels of total EMR2 protein.
  • Tested applications

    Suitable for: WB, ELISAmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide derived from an internal region of human EMR2

  • Positive control

    • HeLa cell extract

Properties

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS

    Without Mg2+ and Ca2+
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    ab75190 was affinity purified from rabbit antiserum by affinity chromatography using epitope specific immunogen.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab75190 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 1/500 - 1/1000. Predicted molecular weight: 91 kDa.
ELISA 1/5000.

Target

  • Function

    Receptor probably involved in cell attachment.
  • Tissue specificity

    Expression is restricted to myeloid cells. Highest expression was found in peripheral blood leukocytes, followed by spleen and lymph nodes, with intermediate to low levels in thymus, bone marrow, fetal liver, placenta, and lung, and no expression in heart, brain, skeletal muscle, kidney, or pancreas. Expression is also detected in monocyte/macrophage and Jurkat cell lines but not in other cell lines tested.
  • Sequence similarities

    Belongs to the G-protein coupled receptor 2 family. LN-TM7 subfamily.
    Contains 5 EGF-like domains.
    Contains 1 GPS domain.
  • Domain

    The GPS domain is necessary, but not sufficient for receptor cleavage, which require the entire extracellular stalk.
    Binding to chondroitin sulfate is mediated by the fourth EGF domain.
  • Post-translational
    modifications

    Autoproteolytically cleaved into 2 subunits, an extracellular alpha subunit and a seven-transmembrane beta subunit.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 312 antibody
    • CD 312 antigen antibody
    • CD312 antibody
    • CD312 antigen antibody
    • Egf like module containing mucin like hormone receptor like 2 antibody
    • Egf like module containing mucin like hormone receptor like sequence 2 antibody
    • EGF like module EMR2 antibody
    • EGF-like module receptor 2 antibody
    • EGF-like module-containing mucin-like hormone receptor-like 2 antibody
    • EMR2 antibody
    • EMR2_HUMAN antibody
    see all

Images

  • All lanes : Anti-EMR2 antibody (ab75190) at 1/500 dilution

    Lane 1 : HeLa cell extract
    Lane 2 : HeLa cell extract with immunising peptide at 10 µg

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 91 kDa
    Observed band size: 91 kDa

References

ab75190 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for your response and for your continued patronage of Abcam. I am glad that you enjoyed the brownies!

When ordering, it is most beneficial to use our website or to call our customer service representatives as they are much more knowledgeable of the ordering system and would be able to effectively resolve any issues should they arise.

Thank you again contacting me and for your kind words. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

I has been my pleasure to help with this. I would like to thank you for your help as well and for your cooperation. The details that you have provided will 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 of this product for ab122999. The order number for this replacement is 1040525.


This product is expected not in stock at the moment but is expected to ship next week. 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. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question

I hope this message finds you well. Thank you for your response, here are my answers to your questions. I have also attached a sample image of the bands we are getting. 1. Denaturing samples - After normalizing to a protein concentration of 1-2 ug/ul, I dilute the samples in 1x SDS and then heat them for 5 minutes at 90-100 degrees C. The samples are stored at -80 C, but heated for 1-2 minutes before use at the same temperature. 2. Each well that you will see has 40 ug protein 3. My loading control is GAPDH, which looks good. 4. My positive controls were mouse spleen and the glioblastoma cell line SF767. Both are known to express EMR2. 5. We used a nitrocellulose membrane 6. Samples were blocked in 5% milk, 3% BSA solution for 90 minutes at room temperature 7. All antibodies were diluted 1:500 and incubated at 4 degrees. 8. Secondary antibodies were goat anti-human, used at 1:2000 dilution incubated at room temperature for 90 minutes 9. I have not run a no primary antibody control blot 10. The wash buffer is 1x TBST 11. I will get the abcam PO number from the lab manager and email you asap. Regarding the attached photo: The lanes are labeled as is the ladder. From the bottom to top the ladder values are (in kD): 37, 50, 75, 100, 150, 250. Our loading control (GAPDH) is at 37 kD as expected, as is the SF767 positive control for EMR2. Mouse spleen runs a little low and we are seeing what appears to be an isoform in the far right lane (GBM 28) at around 80 kD, although this is an isoform that according to the immunogen sequence provided. I am seeing the same isoform (approx 80 kD) more in other blots, but this is the blot that is most readily available for email. Thank you for all of your help and kind consideration. I look forward to hearing from you.

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Answer

Thank you for sending me this further information. It has helped tremendously in understanding the issues that you have been facing.

I agree with your assessment that these bands are either non-specific or isoforms 4 and 5. As we cannot be sure which and I can see no futher suggestions that I might make which you have not already tried. If you have purchase this antibody wuithin the last six months, it will be covered by our Abpromise guarantee. In which case I mayoffer a free of charge replacement for this product.

This issue may be specific to this vial or lot in wehich case I can offer you a new vial of ab75190. I beleive that I have already also communicated the immunogen sequence to you so should you decide to perform an immungen blocking procedure that peptide could easliy be created.

As we had discussed earlier ab122999should recognize isoforms 1, 2, 3 and 4, as there is 100% homology while there is only 76% homology to isoform 5. However, the immungen for the product is published should it be neccessary to create a blocking peptide as well.

I look forward to hearing from you, please be sure to send the Abcam order confirmation number or PO number used. I've included a pdf file overviewing immunogen blocking procedures. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.







I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

I am sorry about the issues you have been having. Thank you for bringing this to our attention. As this product should not be recognizing the other forms of EMR2 it appears that you may be experiencing non specific background issues. I may be able to help you reduce or eliminate these issues if yopu could send me the details of your protocol and any images you may have. I've put a number of questions below which will help me to get the clearest possible picture of the issue you are facing. Please forgive me if we have spoken about some of these before as my notes are rather light in regard to whether we had spoken about your protocols.

I see that you have been working with Human tissues, could you tell me the lysis buffer that you have used?

Have you reduced and denatured your samples? Which method/buffers did you use?

How much total protein did you use per well?

Did you load a loading control? Which LC did you use? What were the results?

Did you load a positive control? Which PC did you use? What were the results?

What membrane type did you transfer to?

What blocking steps did you use?

What dilutions of this product did you use? How long and at what temperature was this antibody incubated?

What was the secondary antibody that you have used? How long and at what temperature was this antibody incubated?

Have you run a no primary antibody control blot? What were the results?

What is the wash buffer that you use?

Could you provide the Abcam order confirmation number or PO number used to purchase this?

Thank you very much for taking the time to answer these questions. Please contact me if you have any questions or concerns.

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Answer

I have had to opportunity to speak with the lab further regarding this. The immunogen sequence for this product is:XXX which spans aaXXX-XXX (SwissProt: Q9UHX3Human). This sequence is within the excellular region and is not in isoforms 4 and 5. Are you experiencing non-specific background bands at these weights? It may help to adjust your protocol slightly to reduce or eliminate these. Some steps that may be taken include changing your blocking solution to 5% BSA if using milk, lowering the concentration of the primary antibody is often helpful as well. The antibody is covered under our Abpromise for six months and is guaranteed to work ib WBon humansamples. If you are experiencing issues with this product and we cannot resolve the issue you are having , then I would be happy to either send a replacement antibody or to process a refund. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you for contacting us.

I have had the opportunity to speak with the labs in regard to your questions about these product.

The immunogen sequence for each product is proprietary but the labs have run alignments and checked homologies with other isoforms. The results are as follows:



ab122999: Based on the peptide immunogen used to generate our anti-EMR2 antibody (ARP61772_P050) and the UniProt EMR2 protein page (http://www.uniprot.org/uniprot/Q9UHX3), this antibody should recognize isoforms 1, 2, 3 and 4, as there is 100% homology. There is only 76% homology to isoform 5 (11 consecutive matches, 3 non-matches on the end of the 14aa immunogen), which is near the threshold of what we would predict for reactivity (80%).



ab108368: We have not tested this antibody for recognition of other isoforms. Based on sequence analysis, the immunogen shares 100% identity with isoforms 2 and 3. It is predicted, but it is not a guarantee without testing.



ab75190: Human EMR2 (CD312) has five isoforms (isoform 1-5). This antibody reacts with isoform 1, 2, and 3. It does not react with isoform 4 and 5. The reaction domain is extracellular domain (on the first topological domain)



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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