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  1. Link

    products/primary-antibodies/tie2-antibody-cl-16-bsa-and-azide-free-ab24859.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)

  • Datasheet
Reviews (13)Q&A (9)References (24)

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Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
  • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
  • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
  • Flow Cytometry - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
  • Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)

Key features and details

  • Mouse monoclonal [Cl. 16] to TIE2 - BSA and Azide free
  • Suitable for: IHC-Fr, WB, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-TIE2 antibody [Cl. 16] - BSA and Azide free
    See all TIE2 primary antibodies
  • Description

    Mouse monoclonal [Cl. 16] to TIE2 - BSA and Azide free
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-Fr, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment corresponding to Human TIE2. Recombinant human soluble extracellular TIE2.
    Database link: Q02763

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    Cl. 16
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factor receptors
    • Cardiovascular
    • Vasculature
    • Endothelium
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Developmental Biology
    • Organogenesis
    • Angiogenesis and vasculogenesis
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers
    • Neuroscience
    • Development

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant human TIE2 protein (ab129039)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab24859 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-Fr (2)
Use at an assay dependent concentration.
WB (1)
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 126 kDa.
Flow Cyt
Use 1-2µg for 106 cells.

We tested in-house with methanol-fixed cells, but this may not be necessary since the antibody recognies an extracellular epitope.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Notes
IHC-Fr
Use at an assay dependent concentration.
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 126 kDa.
Flow Cyt
Use 1-2µg for 106 cells.

We tested in-house with methanol-fixed cells, but this may not be necessary since the antibody recognies an extracellular epitope.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Tyrosine-protein kinase that acts as cell-surface receptor for ANGPT1, ANGPT2 and ANGPT4 and regulates angiogenesis, endothelial cell survival, proliferation, migration, adhesion and cell spreading, reorganization of the actin cytoskeleton, but also maintenance of vascular quiescence. Has anti-inflammatory effects by preventing the leakage of proinflammatory plasma proteins and leukocytes from blood vessels. Required for normal angiogenesis and heart development during embryogenesis. Required for post-natal hematopoiesis. After birth, activates or inhibits angiogenesis, depending on the context. Inhibits angiogenesis and promotes vascular stability in quiescent vessels, where endothelial cells have tight contacts. In quiescent vessels, ANGPT1 oligomers recruit TEK to cell-cell contacts, forming complexes with TEK molecules from adjoining cells, and this leads to preferential activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascades. In migrating endothelial cells that lack cell-cell adhesions, ANGT1 recruits TEK to contacts with the extracellular matrix, leading to the formation of focal adhesion complexes, activation of PTK2/FAK and of the downstream kinases MAPK1/ERK2 and MAPK3/ERK1, and ultimately to the stimulation of sprouting angiogenesis. ANGPT1 signaling triggers receptor dimerization and autophosphorylation at specific tyrosine residues that then serve as binding sites for scaffold proteins and effectors. Signaling is modulated by ANGPT2 that has lower affinity for TEK, can promote TEK autophosphorylation in the absence of ANGPT1, but inhibits ANGPT1-mediated signaling by competing for the same binding site. Signaling is also modulated by formation of heterodimers with TIE1, and by proteolytic processing that gives rise to a soluble TEK extracellular domain. The soluble extracellular domain modulates signaling by functioning as decoy receptor for angiopoietins. TEK phosphorylates DOK2, GRB7, GRB14, PIK3R1; SHC1 and TIE1.
  • Tissue specificity

    Detected in umbilical vein endothelial cells. Proteolytic processing gives rise to a soluble extracellular domain that is detected in blood plasma (at protein level). Predominantly expressed in endothelial cells and their progenitors, the angioblasts. Has been directly found in placenta and lung, with a lower level in umbilical vein endothelial cells, brain and kidney.
  • Involvement in disease

    Dominantly inherited venous malformations
    May play a role in a range of diseases with a vascular component, including neovascularization of tumors, psoriasis and inflammation.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. Tie subfamily.
    Contains 3 EGF-like domains.
    Contains 3 fibronectin type-III domains.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.
  • Domain

    The soluble extracellular domain is functionally active in angiopoietin binding and can modulate the activity of the membrane-bound form by competing for angiopoietins.
  • Post-translational
    modifications

    Proteolytic processing leads to the shedding of the extracellular domain (soluble TIE-2 alias sTIE-2).
    Autophosphorylated on tyrosine residues in response to ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Autophosphorylation occurs in a sequential manner, where Tyr-992 in the kinase activation loop is phosphorylated first, followed by autophosphorylation at Tyr-1108 and at additional tyrosine residues. ANGPT1-induced phosphorylation is impaired during hypoxia, due to increased expression of ANGPT2. Phosphorylation is important for interaction with GRB14, PIK3R1 and PTPN11. Phosphorylation at Tyr-1102 is important for interaction with SHC1, GRB2 and GRB7. Phosphorylation at Tyr-1108 is important for interaction with DOK2 and for coupling to downstream signal transduction pathways in endothelial cells. Dephosphorylated by PTPRB.
    Ubiquitinated. The phosphorylated receptor is ubiquitinated and internalized, leading to its degradation.
  • Cellular localization

    Cell membrane. Cell junction. Cell junction, focal adhesion. Cytoplasm, cytoskeleton. Secreted. Recruited to cell-cell contacts in quiescent endothelial cells. Colocalizes with the actin cytoskeleton and at actin stress fibers during cell spreading. Recruited to the lower surface of migrating cells, especially the rear end of the cell. Proteolytic processing gives rise to a soluble extracellular domain that is secreted.
  • Target information above from: UniProt accession Q02763 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7010 Human
    • Omim: 600221 Human
    • SwissProt: Q02763 Human
    • Unigene: 89640 Human
    • Alternative names

      • Angiopoietin 1 receptor antibody
      • Angiopoietin-1 receptor antibody
      • CD202b antibody
      • CD202b antigen antibody
      • Endothelial tyrosine kinase antibody
      • Endothelium specific receptor tyrosine kinase 2 antibody
      • hTIE 2 antibody
      • hTIE2 antibody
      • Hyk antibody
      • p140 TEK antibody
      • Soluble TIE2 variant 1 antibody
      • Soluble TIE2 variant 2 antibody
      • Tek antibody
      • tek tyrosine kinase antibody
      • TEK tyrosine kinase endothelial antibody
      • tek tyrosine kinase, endothelial antibody
      • TIE 2 antibody
      • TIE2 antibody
      • TIE2_HUMAN antibody
      • Tunica interna endothelial cell kinase antibody
      • Tyrosine kinase with Ig and EGF homology domains 2 antibody
      • Tyrosine kinase with Ig and EGF homology domains-2 antibody
      • Tyrosine protein kinase receptor TEK antibody
      • Tyrosine protein kinase receptor TIE 2 antibody
      • Tyrosine-protein kinase receptor TEK antibody
      • Tyrosine-protein kinase receptor TIE-2 antibody
      • Venous malformations multiple cutaneous and mucosal antibody
      • VMCM 1 antibody
      • VMCM antibody
      • VMCM1 antibody
      see all

    Images

    • Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
    • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      ab24859 staining TIE2 in Human spleen by Immunohistochemistry (Frozen sections).
    • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)This image is courtesy of an anonymous Abreview
      Immunohistochemical analysis of Human brain tissue, staining TIE2 with ab24859.

      Tissue was fixed with paraformaldehyde, permeabilized with 0.25 Triton X-100 and blocked with 2.5% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200 in 2.5% horse serum) for 18 hours at 4°C. An HRP-conjugated horse anti-rabbit polyclonal IgG was used as the secondary antibody.

      See Abreview

    • Flow Cytometry - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Flow Cytometry - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Overlay histogram showing JEG-3 cells stained with ab24859 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24859, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in JEG-3 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

      Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
    • Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)
      All lanes : Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (ab24859)

      Lane 1 : HUVECs left untreated
      Lane 2 : HUVECs stimulated for 3 hours with PMA at 25 ng/ml
      Lane 3 : HUVECs stimulated for 6 hours with PMA at 25 ng/ml
      Lane 4 : HUVECs stimulated for 9 hours with PMA at 25 ng/ml
      Lane 5 : HUVECs stimulated for 24 hours with PMA at 25 ng/ml

      Predicted band size: 126 kDa



      Samples were immunoprecipitated with another TIE2 antibody.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (24)

    Publishing research using ab24859? Please let us know so that we can cite the reference in this datasheet.

    ab24859 has been referenced in 24 publications.

    • Gebara N  et al. Single extracellular vesicle analysis in human amniotic fluid shows evidence of phenotype alterations in preeclampsia. J Extracell Vesicles 11:e12217 (2022). PubMed: 35582873
    • Shi C  et al. NEAT1 promotes the repair of abdominal aortic aneurysms of endothelial progenitor cells via regulating miR-204-5p/Ang-1. Am J Transl Res 13:2111-2126 (2021). PubMed: 34017378
    • Pushpakumar S  et al. Exogenous hydrogen sulfide and miR-21 antagonism attenuates macrophage-mediated inflammation in ischemia reperfusion injury of the aged kidney. Geroscience 43:1349-1367 (2021). PubMed: 33433751
    • Hassanpour M  et al. Resveratrol reduced the detrimental effects of malondialdehyde on human endothelial cells. J Cardiovasc Thorac Res 13:131-140 (2021). PubMed: 34326967
    • Lee YN  et al. Deferoxamine accelerates endothelial progenitor cell senescence and compromises angiogenesis. Aging (Albany NY) 13:21364-21384 (2021). PubMed: 34508614
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-9 of 9 Q&A

    Question

    Hello, I have a couple of questions about your Tie2 antibodies (ab24859 and ab47183). First, do you know the binding sites for these antibodies on the Tie2 receptor? Do they show any antagonistic activity - we need to preserve angiopoietin binding and signalling. Second, why is ab24859 more expensive than ab47183? We are doing single particle fluorescence tracking on live cells. Thank you very much

    Read More

    Abcam community

    Verified customer

    Asked on Sep 18 2012

    Answer

    Thank you for your patience in this matter and I do apologize for the delay.

    Unfortunately we do not have any information about any agonistic activity of our anti-human TIE-2, ab24859.

    I am sorry about that I cannot provide any more information for you, but if there is anything else I can help you with, please let me know.

    Read More

    Abcam Scientific Support

    Answered on Sep 18 2012

    Question

    Hello, I have a couple of questions about your Tie2 antibodies (ab24859 and ab47183). First, do you know the binding sites for these antibodies on the Tie2 receptor? Do they show any antagonistic activity - we need to preserve angiopoietin binding and signalling. Second, why is ab24859 more expensive than ab47183? We are doing single particle fluorescence tracking on live cells. Thank you very much

    Read More

    Abcam community

    Verified customer

    Asked on Aug 16 2012

    Answer

    Thank you for contacting to Abcam.


    Please find the answers to your questions below:

    Question 1: I was informed by the lab that epitope mapping was carried out on this antibody a while back, and they are looking through their notes to see if it was successful or not. If it was, I will pass on that information when it is available.

    Question 2: The antibody did not show any antagonistic activity.

    If there is anything else I can help you with, please let me know.

    Read More

    Abcam Scientific Support

    Answered on Aug 16 2012

    Question

    Thank you for the assistance. Meanwhile, the customer decided to receive a credit. Please issue Zotal a credit note.Thanks again for your cooperation.Best Regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 22 2012

    Answer

    Thank you for contacting us.
    Your credit note ID is XXXXXXX.
    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:
    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.
    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.
    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.
    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.
    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Abcam Scientific Support

    Answered on May 22 2012

    Question

    The customer followed your suggestion of heating sample at 100C for 15 minutes is little longer. Please try 5 minutes at 95-100C for best results.
    They heated 5 min in 95C, with the same result.
    In the attached files you will see the results.The exposure for 70 sec resulted in no band in the lane with the recombinant protein
    lanes from right to left:
    recombinant tie
    jeg 3 lysates
    mda-mb231 lysates
    lcc lysates
    u87 lysates
    The lower part of the same membrane shows the result of anti alpha tubulin, 10 sec exposure. You can observe a specific band.The customer would like to replace this antibody with another anti tie-2 that reacts with mouse and human for WB and FACS (if FACS is not an option, so the best choice for WB from your experience).
    Ordering details:
    Po: 88805
    Invoice: 1313995
    Thanks in advance for your assistance.
    Have a nice week.

    Read More

    Abcam community

    Verified customer

    Asked on May 21 2012

    Answer

    I am sorry to hear that the results did not improve.
    We have same lot in stock so would you like to try the same lot? Alternatively I can provide ab71712 which was tested in mouse and is predicted to react with human species. This antibody hasn't been tested in Flow Cytormetry however this does not mean it can't be used.
    Let me know how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on May 21 2012

    Question

    Product code: 24859
    Lot number: GR71670-1
    Inquiry: Please read below the details of customer's complaint: Antibody code: ab24859 Batch number: lot # GR71670-1 Antibody storage conditions (temperature/reconstitution etc): -80oC in 15 microliter aliquotes Description of the problem (high background, wrong band size, more bands, no band etc.): high background + wrong band size, more bands Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.): cell and tumor lysates and recombinant protein (#1 in gel) Sample preparation (Buffer/Protease inhibitors/Heating sample etc.): RIPA and protease inhibitor with reducing agent and sample buffer, heating- 15 min (95 oC) Amount of protein loaded: 30 microgramm Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.): 10% criterion XT gel- BioRad Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Transfer: 100 Volt constant 1:05h on ice Blocking: 30 min, RT, shaking, 3%BSA in TBST Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Anti tie-2 antibody 1:1000 dilution in 3%BSA in TBST (picture #1) and 1:330 dilution in 3%BSA in TBST (picture #2) – O/N, 4 oC, shaking Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): Goat anti mouse, 1:10000 dilutions in 3%BSA in TBST, 1h, RT, shaking Detection method (ECL, ECLPlus etc.): ECL Positive and negative controls used (please specify): positive- recombinant protein tie-2 protein (Fc chimera)- ab54435 OPTIMIZATION ATTEMPTS (PROBLEM SOLVING) How many times have you tried the Western? 1- 1:1000 dilution and 1-1:330 dilution Have you run a "No Primary" control? Do you obtain the same results every time? e.g. are the background bands always in the same place? yes Please advise. Thanks in advance for your assistance and reply.

    Read More

    Abcam community

    Verified customer

    Asked on May 08 2012

    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.
    The protocol seems fine to me however heating sample at 100C for 15 minutes is little longer. Please try 5 minutes at 95-100C for best results.
    I hope the result will improve after changing this step; if not, I will then be happy to provide a free of charge replacement antibody or a same antibody but different lot.

    Read More

    Abcam Scientific Support

    Answered on May 08 2012

    Question

    Cross reactivity with mouse IHC-Fr

    Read More

    Abcam community

    Verified customer

    Asked on Feb 24 2012

    Answer

    In the publication PMID 19253934 the antibody was indeed used in IHC-Fr however with human samples so I have decide to keep the IHC-Fr application.

    Just to let you know the antibody is still eligible for mouse IHC-FR discount.

    Read More

    Abcam Scientific Support

    Answered on Feb 24 2012

    Question

    Cross reactivity with mouse IHC-Fr

    Read More

    Abcam community

    Verified customer

    Asked on Feb 24 2012

    Answer

    Thank you for contacting us.

    I have checked the Immunogen sequence homology between human and mouse; the extracellular domain of these species is quite similar, which indicates the antibody will cross react.

    http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20120224-163041-0072-66196796-pg

    http://www.ebi.ac.uk/Tools/services/web/toolresult.ebi?jobId=clustalw2-I20120224-163041-0072-66196796-pg&tool=clustalw2&analysis=summary

    In the publication PMID 20622121, the antibody is indeed used in immunohistochemistry with some nice results fig 6. The protocol however is not very clear, it seems they have used this antibody in paraffin embedded sections because antigen retrieval can only be done with IHC-P not IHC-Fr. I therefore can provide you a discount code if you would like to test the antibody in IHC-Fr. Please click the below link to know more about the collaboration discount;

    https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount.

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

    Read More

    Abcam Scientific Support

    Answered on Feb 24 2012

    Question

    Hi Jhon.



    There is the researcher results.



    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 08 2012

    Answer

    Thank you for sending that protocol information.

    If I have read the information correctly, then there are a few protocol variations that may work:

    1 - Did the customer only try a 1/5000 primary antibody dilution? If so, I strongly suggest that they try range of dilutions from 1/500-1/2000, to see if they can find the concentration that will work for them.

    2 - According to the protocol, they loaded 5ug of protein on the gel. Did they try increasing the amount of protein, to say 20ug?

    3 - How long did they incubate the primary antibody for? We recommend overnight at 4C.

    4 - Did they try using 5% milk as a blocking reagent instead of BSA?

    If the customer has not tried the protocol variations above, then I would strongly suggest that they do. If they have already tried those variations, then I would be happy to process a refund for this antibody, as it is not working as we guarantee.

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Feb 09 2012

    Question

    Why do cells have to be methanol fixed for Flow Cytometry?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 15 2011

    Answer

    Thank you for your inquiry. I heard back from the testing laboratory that methanol fixation is not necessary for FC since this antibody recognizes an extracellular epitope. However, our in-house testing followed a methanol fixation protocol and this worked well. I hope this information helps. Please contact us with any other questions.

    Read More

    Abcam Scientific Support

    Answered on Aug 15 2011

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