Overview

  • Product name
  • Description
    Rabbit polyclonal to Ephrin B3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Cow, Cat, Dog, Pig, Macaque monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Ephrin B3.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Human brain; Mouse brain; Rat brain; Mouse forebrain; Rat forebrain; Mouse Embryonic brain E10; Mouse embryonic brain E14.

Properties

Applications

Our Abpromise guarantee covers the use of ab101699 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 36 kDa).
IHC Use at an assay dependent concentration. PubMed: 25309991

Target

  • Function
    May play a pivotal role in forebrain function. Binds to, and induce the collapse of, commissural axons/growth cones in vitro. May play a role in constraining the orientation of longitudinally projecting axons.
  • Tissue specificity
    Highly expressed in brain; expressed in embryonic floor plate, roof plate and hindbrain segments.
  • Sequence similarities
    Belongs to the ephrin family.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • EFL 6 antibody
    • EFL6 antibody
    • EFNB 3 antibody
    • Efnb3 antibody
    • EFNB3_HUMAN antibody
    • EPH related receptor transmembrane ligand ELK L3 antibody
    • EPH related receptor transmembrane ligand ELKL3 antibody
    • EPH related receptor tyrosine kinase ligand 8 antibody
    • EPH-related receptor transmembrane ligand ELK-L3 antibody
    • EPH-related receptor tyrosine kinase ligand 8 antibody
    • Ephrin B3 antibody
    • Ephrin-B3 antibody
    • EphrinB3 antibody
    • EPLG 8 antibody
    • EPLG8 antibody
    • LERK 8 antibody
    • LERK-8 antibody
    • LERK8 antibody
    see all

Images

  • All lanes : Anti-Ephrin B3 antibody (ab101699) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Brain (Rat) Tissue Lysate
    Lane 4 : Forebrain (Mouse) Tissue Lysate
    Lane 5 : Rat Forebrain
    Lane 6 : E10 Mouse Embryo Brain Tissue Lysate
    Lane 7 : E14 Mouse Embryo Brain Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 90 seconds


    Ephrin B3 contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ICC/IF image of ab101699 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab101699, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Efazat G  et al. Ephrin B3 interacts with multiple EphA receptors and drives migration and invasion in non-small cell lung cancer. Oncotarget 7:60332-60347 (2016). Read more (PubMed: 27533087) »
  • Qu B  et al. Testing stem cell therapy in a rat model of inflammatory bowel disease: role of bone marrow stem cells and stem cell factor in mucosal regeneration. PLoS One 9:e107891 (2014). IHC ; Rat . Read more (PubMed: 25309991) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Flow Cytometry
Sample
Mouse Cell (small cell lung cancer cell line 16T)
Permeabilization
No
Gating Strategy
single cells with FSC and SSC, live cells with low DAPI, positive/negative cells with PI filter set, anti-CD24 as positive control
Specification
small cell lung cancer cell line 16T
Preparation
Cell harvesting/tissue preparation method: Cell harvesting: cells grow as floating spheres. Collect by spinning, dissociate spheres with TrypLE, single cell through 40um cell strainer
Sample buffer: PBS + 10% FBS
Fixation
none

Abcam user community

Verified customer

Submitted Jul 26 2018

Answer

DISCOUNT CODES: 100%ABR-E7TFV for testing ab89461 in Zebrafish 100%ABR-E7TF3 for testing ab67168 in Zebrafish 100%ABR-E7TFM for testing ab101699 in Zebrafish Expiration date for all the three codes: 22. February 2012 I am very pleased to hear you would like to accept our offer and test all these three antibodies in Zebrafish. The codes will give you each 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for Zebrafish and include the respective code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your inquiry. Indeed, it is difficult to find antibodies crossreactive with Zebrafish. I don't know whether I have already told you about our list of Zebrafish crossreactive antibodies? It can be found under the following link: www.abcam.com/zebrafish In regards to choosing the most probably crossreacting antibody for your three targets of interest, please find my comments here below. As for the previous antibody, please do not hesitate to ask for the discount code for testing these antibodies on Zebrafish (www.abcam.com/collaborationdiscount) I would like also to recommend, to chose antibodies which have been tested in the application or at least a similar application to which you will be using. Antibodies against Neurogenin 1: The ab89461 has a good alignment score and it seems likely that crossreactivity could happen. The score is 68% http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-113055-0569-24134208-pg https://www.abcam.com/index.html?datasheet=89461 Also the ab117847 has reasonable chances to detect the Zebrafish protein, as the full length human protein has been used as an immunogen and the score is 47%. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-105747-0570-80392552-pg https://www.abcam.com/index.html?datasheet=117847 The two other antibodies, ab66498 and ab71818, are generated against peptides from N and C term, and as the homology there is not very good with the Zebrafish protein, I would recommend to test the other two antibodies first. NeuroD4 The immunogen of ab90484 has stretch conserved and I think it could work therefore. The score is 50% and the alignment can be found here: http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-110656-0883-28043163-pg https://www.abcam.com/index.html?datasheet=90484 The ab67168 has been produced with the full length human protein and the overall score is with 55% also relatively ok. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-110901-0500-99792871-pg https://www.abcam.com/index.html?datasheet=67168 Ephrin B3 The overall protein alignment between the human and Zebrafish protein is relatively good, with a score of 54%.http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-111305-0838-36391612-pg The ab101699 has a reasonable good alignement and I could recommend to test this antibody therefore. The alignment is ********:::***: with a score of 73% http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-111510-0668-3570069-pg https://www.abcam.com/index.html?datasheet=101699 Of the other antibodies, I can also recommend to test either the ab28389 or the ab89474, as they have been produced with relatively long sequences and possibility of crossreactivity is therefore good as well. https://www.abcam.com/index.html?datasheet=28389 https://www.abcam.com/index.html?datasheet=89474 I would not recommend the ab53063 and for the ab103001 and ab106344 I do not know the immunogen sequences. If however these antibodies would interest you also in regards to their tested applications, I will be happy to ask the laboratory for prediction of the crossreaction. I hope this information is helpful. Please do not hesitate to contact me for the discount codes or for further information.

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