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Cardiovascular Angiogenesis Ephrins Eph Receptors

Anti-Eph receptor A4/SEK抗体(ab5396)

  • Datasheet
Reviews (4)Q&A (6)References (4)

Product price, shipping and contact information

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Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
  • Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor A4/SEK antibody (ab5396)

Key features and details

  • Rabbit polyclonal to Eph receptor A4/SEK
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Eph receptor A4/SEK抗体
    参阅全部 Eph receptor A4/SEK 一抗
  • 描述

    兔多克隆抗体to Eph receptor A4/SEK
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, IHC-Pmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human Eph receptor A4/SEK aa 875-904 (C terminal) conjugated to keyhole limpet haemocyanin.

  • 常规说明

    Protein kinases are enzymes that transfer a phosphate group from a phosphate donor, generally the g phosphate of ATP, onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. With more than 500 gene products, the protein kinase family is one of the largest families of proteins in eukaryotes. The family has been classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. The tyrosine kinase (TK) group is mainly involved in the regulation of cell-cell interactions such as differentiation, adhesion, motility and death. There are currently about 90 TK genes sequenced, 58 are of receptor protein TK (e.g. EGFR, EPH, FGFR, PDGFR, TRK, and VEGFR families), and 32 of cytosolic TK (e.g. ABL, FAK, JAK, and SRC families).

     This product was previously labelled as Eph receptor A4

     

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度

    Protein G purified
  • 纯化说明

    This antibody is purified through a protein G column and eluted out with both high and low pH buffers and neutralized immediately after elution then followed by dialysis against PBS.
  • Primary antibody说明

    Protein kinases are enzymes that transfer a phosphate group from a phosphate donor, generally the g phosphate of ATP, onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. With more than 500 gene products, the protein kinase family is one of the largest families of proteins in eukaryotes. The family has been classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. The tyrosine kinase (TK) group is mainly involved in the regulation of cell-cell interactions such as differentiation, adhesion, motility and death. There are currently about 90 TK genes sequenced, 58 are of receptor protein TK (e.g. EGFR, EPH, FGFR, PDGFR, TRK, and VEGFR families), and 32 of cytosolic TK (e.g. ABL, FAK, JAK, and SRC families).
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Cardiovascular
    • Angiogenesis
    • Ephrins
    • Eph Receptors
    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Kinases
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Growth Cone
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Neuroscience
    • Neurology process
    • Growth and Development
    • Axonal Guidance Proteins
    • Neuroscience
    • Neurotransmission
    • Receptors / Channels
    • Tyrosine Kinase Receptors

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Eph receptor A4/SEK protein (ab84575)

应用

Our Abpromise guarantee covers the use of ab5396 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 108 kDa (predicted molecular weight: 118 kDa).

We recommend using CHAPS lysis buffer rather than RIPA, Invitrogen NuPage gel system, blotting on PVDF membrane, blocking 30' in 10% Fetal Bovine Serum, 3% BSA, 0.3% Tween 20 in 1X PBS - please see Abreview for further information (published on 24th July, 2006).

IHC-P 1/25 - 1/100.

靶标

  • 功能

    Receptor for members of the ephrin-A family. Binds to ephrin-A1, -A4 and -A5. Binds more poorly to ephrin-A2 and -A3. May play a role in a signal transduction process involved in hindbrain pattern formation.
  • 组织特异性

    Ubiquitous.
  • 序列相似性

    Belongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 1 protein kinase domain.
    Contains 1 SAM (sterile alpha motif) domain.
  • 结构域

    The protein kinase domain mediates interaction with NGEF/ephexin-1.
  • 细胞定位

    Membrane.
  • Target information above from: UniProt accession P54764 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 2043 Human
    • Entrez Gene: 13838 Mouse
    • Entrez Gene: 316539 Rat
    • Omim: 602188 Human
    • SwissProt: P54764 Human
    • SwissProt: Q03137 Mouse
    • Unigene: 371218 Human
    • Unigene: 400747 Mouse
    • 别名

      • Cek 8 antibody
      • CEK8 antibody
      • EK8 antibody
      • eph receptor a4 antibody
      • EPH-like kinase 8 antibody
      • EPHA4 antibody
      • EPHA4_HUMAN antibody
      • Ephrin type-A receptor 4 antibody
      • HEK 8 antibody
      • hEK8 antibody
      • Receptor protein-tyrosine kinase HEK8 antibody
      • Sek 1 antibody
      • SEK antibody
      • TYRO 1 protein tyrosine kinase antibody
      • TYRO1 antibody
      • Tyrosine-protein kinase receptor SEK antibody
      • Tyrosine-protein kinase TYRO1 antibody
      see all

    图片

    • Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
      Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
      All lanes : Anti-Eph receptor A4/SEK antibody (ab5396) at 1/1000 dilution

      Lane 1 : HeLa Cell Lysate
      Lane 2 : NCI-H460 Cell Lysate
      Lane 3 : Mouse NIH3T3 Cell Lysate
      Lane 4 : Human ovary tissue Lysate

      Lysates/proteins at 35 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG H+L HRP conjugated at 1/10000 dilution

      Predicted band size: 118 kDa



      Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
      Western blot - Anti-Eph receptor A4/SEK antibody (ab5396)
      Anti-Eph receptor A4/SEK antibody (ab5396) at 1/1000 dilution + NCI-H460 whole cell lysate at 35 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/10000 dilution

      Predicted band size: 118 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor A4/SEK antibody (ab5396)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor A4/SEK antibody (ab5396)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling Eph receptor A4/SEK with ab5396 at dilution 1/25. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 37°C; heat mediated antigen retrieval was performed using a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hour at 37°C. A Peroxidase-conjugated Goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.

    实验方案

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet
  • 文献 (4)

    发表研究结果有使用 ab5396?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab5396 被引用在 4 文献中.

    • Feng L  et al. EphA4 may contribute to microvessel remodeling in the hippocampal CA1 and CA3 areas in a mouse model of temporal lobe epilepsy. Mol Med Rep 15:37-46 (2017). PubMed: 27959424
    • Todd KL  et al. EphA4 Regulates Neuroblast and Astrocyte Organization in a Neurogenic Niche. J Neurosci 37:3331-3341 (2017). ICC/IF ; Mouse . PubMed: 28258169
    • Fan R  et al. Critical role of EphA4 in early brain injury after subarachnoid hemorrhage in rat. Exp Neurol 296:41-48 (2017). PubMed: 28698029
    • Wu LY  et al. Connexin 43 stabilizes astrocytes in a stroke-like milieu to facilitate neuronal recovery. Acta Pharmacol Sin 36:928-38 (2015). PubMed: 26095039

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 10 Abreviews or Q&A

    IHC - Wholemount abreview for Anti-Eph receptor A4/SEK antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Common marmoset Tissue (Adult Brain)
    Specification
    Adult Brain
    Read More

    Dr. 煜欽 林

    Verified customer

    提交于 Jun 05 2013

    Question

    Have you tested it on western blot so you know that it is the correct size?

    Kind Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 14 2013

    Answer

    The new lot has been tested in WB so it will be OK.

    I have requested image from laboratory, which I forward to you soon.

    Read More

    Abcam Scientific Support

    回复于 Jan 14 2013

    Question

    We would like to try a different lot of EPHA4 antibody that you can guarantee is ok. We have recently bought another antibody (ROBLD3) from you which we put a lot of effort in that turned out to be against another protein.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 14 2013

    Answer

    Thank you very much for having patience.

    We have now acquired new lot of this antibody. I will inform you soon with the shipment date.

    Please let me know if you have any question.

    Read More

    Abcam Scientific Support

    回复于 Jan 14 2013

    Question

    We would like to try a different lot of EPHA4 antibody that you can guarantee is ok. We have recently bought another antibody (ROBLD3) from you which we put a lot of effort in that turned out to be against another protein.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 04 2013

    Answer

    Thank you for your email.

    I have been in contact with laboratory for acquiring new lot. I will send you a confirmation once we ship it.

    Many thanks for having patience. Please do not hesitate to contact me if you have any question.

    Read More

    Abcam Scientific Support

    回复于 Jan 04 2013

    Question

    Here are the answers to the questions. Hope they can clarify.

    Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 24 2012

    Answer

    Dr. Tanya is on holiday until 2nd January so I am dealing with her inquires.

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

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

    I apologize for the inconvenience and am pleased to offer you a free of charge replacement from different lot.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    PS: Please provide the purchase order number which will help us to send you the replacement at same address if you are interested in trying a different lot.

    Read More

    Abcam Scientific Support

    回复于 Dec 24 2012

    Question

    Hi,



    Here is an image of IHC and western blot for EPHA4 and some information on the IHC protocol.



    IHC: We often obtain nuclear staining. Have you found that also?



    Western: The size is approx 90-95 (three different cell lines) but the predicted size is 108kD.



    1) Abcam product code ab 5396



    2) Abcam order reference number or product batch number GR223521



    3) Description of the problem attached IHC image. We often observe nuclear staining. Have you experienced that?



    4) Sample preparation:

    Species

    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other:

    Sample preparation

    Positive control

    Negative control



    5) Fixation step

    Yes/No

    If yes: Fixative agent and concentration

    Fixation time

    Fixation temperature



    6) Antigen retrieval method reveal in in a decloaking chamber according to the manufacturer’s instructions (Biocare, Concord, CA, USA)



    7) Permeabilization method:

    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

    Permeabilizing agent and concentration:





    8) Blocking agent (eg BSA, serum…): 2.5 % horse serum

    Concentration

    Blocking time 20 min

    Blocking temperature room temperature



    9) Endogenous peroxidases blocked?yes

    Endogenous biotins blocked?



    10) Primary antibody (If more than one was used, describe in “additional notes”) :EPHA4

    Concentration or dilution 1:100

    Diluent buffer

    Incubation time



    11) Secondary antibody: incubation using streptavidin/peroxidase complex was performed according to the manufacturer’s manual (Vectastain Universal Quick Kit, Vector Laboratories Inc., Burlingame, CA, USA)

    Species:

    Reacts against:

    Concentration or dilution

    Diluent buffer

    Incubation time

    Fluorochrome or enzyme conjugate



    12) Washing after primary and secondary antibodies:yes

    Buffer TBS

    Number of washes 3



    13) Detection method incubation using streptavidin/peroxidase complex was performed according to the manufacturer’s manual (Vectastain Universal Quick Kit, Vector Laboratories Inc., Burlingame, CA, and development step with 3,3’́-diaminobenzidine



    14) How many times have you run this staining?

    Do you obtain the same results every time?

    What steps have you altered to try and optimize the use of this antibody?



    Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.





    I hope that you can help to clarify this.



    Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 20 2012

    Answer

    Thank you for your enquiry regarding ab5396 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

    Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem. I need to deal with the 2 applications separately and appreciate some further details:

    IMMUNOSTAINING:

    1) Sample preparation: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other

    Question: Could you please specify the species, cell or tissue types used? This information is crucial for us for troubleshooting and it is not provided in this and your previous e-mail.

    2) If yes: Fixative agent and concentration

    Question: What exactly used for fixing the sample? Could you please confirm? Aldehyde or alkohol-based?

    3) Antigen retrieval method reveal in in a decloaking chamber according to the manufacturer’s instructions (Biocare, Concord, CA, USA)

    Question: Did you apply heat or enzyme-based antigen retrial and any optimization steps were performed?

    4) Blocking agent (eg BSA, serum…): 2.5 % horse serum, Blocking time 20 min

    Question: Have you tried 10% serum or 5% BSA for 1 hr at RT?

    WESTERN BLOT:

    Questions:

    - What lysis buffer was used for sample preparation i.e. any proteinsea inhibitors included?

    - How the non-specific bindings were blocked?

    - How much protein was loaded?

    - Has the detection system been checked or no primary antibody run?

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

    Read More

    Abcam Scientific Support

    回复于 Dec 20 2012

    Question

    Hi,

    We have been testing EPHA4 ab abcam5396 for IHC and observed some nuclear staining. Have you seen that with this antibody? What antigen retrieval did you use?

    We have also done immunoblotting and get a product between 90-95 kD (predicted 118 kD). Have you seen this before?

    We feel unsure whether it is an EPHA4 antibody. How well have you tested it?

    Kind Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 12 2012

    Answer

    I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

    I can confirm that ab5396 has been tested for both Western blot and for Immunohistochemistry.

    In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

    As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

    Could you please provide some further details of the protocol used and complete the following forms WB and IHC (attached as word documents). It would be much appreciated if you could attach an image to the response.

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

    Read More

    Abcam Scientific Support

    回复于 Dec 12 2012

    Western blot abreview for Anti-Eph receptor A4/SEK antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain C57Bl/6 mice)
    Loading amount
    30 µg
    Specification
    Brain C57Bl/6 mice
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3%
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 23 2006

    Western blot abreview for Anti-Eph receptor A4 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (P0 RAT Spinal Cord)
    Loading amount
    30 µg
    Specification
    P0 RAT Spinal Cord
    Blocking step
    Other as blocking agent for 30 minute(s) · Concentration: 3%BSA, 10%
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 14 2006

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Eph receptor A4/SEK antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Cerebellar granule cells)
    Specification
    Cerebellar granule cells
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% goat se
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 09 2006

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