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使用敲除细胞株进行验证重组RabMAb

重组Anti-MERTK抗体[Y323] (ab52968)

Reviews (4)Q&A (6)References (61)
Western blot - Anti-MERTK antibody [Y323] (ab52968)
  • Western blot - Anti-MERTK antibody [Y323] (ab52968)
  • Western blot - Anti-MERTK antibody [Y323] (ab52968)
  • Western blot - Anti-MERTK antibody [Y323] (ab52968)
  • Immunoprecipitation - Anti-MERTK antibody [Y323] (ab52968)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)
  • Immunohistochemistry (Frozen sections) - Anti-MERTK antibody [Y323] (ab52968)
  • Western blot - Anti-MERTK antibody [Y323] (ab52968)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)
  • Anti-MERTK antibody [Y323] (ab52968)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y323] to MERTK
  • Suitable for: WB, IP, IHC-P, IHC-Fr
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Carrier Free HRP

概述

  • 产品名称

    Anti-MERTK抗体[Y323]
    参阅全部 MERTK 一抗

  • 描述

    兔单克隆抗体[Y323] to MERTK

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, IHC-P, IHC-Frmore details
    不适用于: Flow Cyt or ICC/IF

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide within Human MERTK aa 1-100 (N terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: A172, HEK-293 and K562 cell lysate. ild-type HAP1 whole cell lysate IHC-P: Human tonsil and lymphoma tissues. IP: MERTK IP in HEK-293 cell lysate. IHC-Fr: Human spleen tissue sections.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab52968于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (1)
1/2000. Detects a band of approximately 180 kDa.

For unpurified use at 1/500.
IP
1/20.

For unpurified use at 1/50.
IHC-P (3)
1/1000 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
IHC-Fr
Use at an assay dependent concentration.
说明
WB
1/2000. Detects a band of approximately 180 kDa.

For unpurified use at 1/500.
IP
1/20.

For unpurified use at 1/50.
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
IHC-Fr
Use at an assay dependent concentration.
应用说明
Is unsuitable for Flow Cyt or ICC/IF.

靶标

  • 功能

    In case of filovirus infection, seems to function as a cell entry factor.

  • 组织特异性

    Not expressed in normal B- and T-lymphocytes but is expressed in numerous neoplastic B- and T-cell lines.

  • 疾病相关

    Defects in MERTK are the cause of retinitis pigmentosa type 38 (RP38) [MIM:613862]. RP38 is a retinal dystrophy belonging to the group of pigmentary retinopathies. Retinitis pigmentosa is characterized by retinal pigment deposits visible on fundus examination and primary loss of rod photoreceptor cells followed by secondary loss of cone photoreceptors. Patients typically have night vision blindness and loss of midperipheral visual field. As their condition progresses, they lose their far peripheral visual field and eventually central vision as well.

  • 序列相似性

    Belongs to the protein kinase superfamily. Tyr protein kinase family. AXL/UFO subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.

  • 细胞定位

    Membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

      • c MER antibody
      • c mer proto oncogene tyrosine kinase antibody
      • c-mer antibody
      • cMER antibody
      • cmer protooncogene tyrosine kinase antibody
      • Eyk antibody
      • MER antibody
      • MER receptor tyrosine kinase antibody
      • MERK antibody
      • MERPEN antibody
      • Mertk antibody
      • MERTK c-mer proto-oncogene tyrosine kinase antibody
      • MERTK_HUMAN antibody
      • MGC133349 antibody
      • nmf12 antibody
      • Nyk antibody
      • Proto oncogene tyrosine protein kinase MER antibody
      • Proto oncogene tyrosine protein kinase MER precursor antibody
      • Proto-oncogene c-Mer antibody
      • Receptor tyrosine kinase MerTK antibody
      • RP38 antibody
      • STK kinase antibody
      • Tyrosine-protein kinase Mer antibody
      see all

    图片

    • Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Western blot - Anti-MERTK antibody [Y323] (ab52968)
      All lanes : Anti-MERTK antibody [Y323] (ab52968) at 1/1000 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : MERTK knockout A549 cell lysate
      Lane 3 : HAP1 cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Observed band size: 160 kDa why is the actual band size different from the predicted?



      Western blot: Anti-MERTK antibody [Y323] (ab52968) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52968 was shown to bind specifically to MERTK. A band was observed at 160 kDa in wild-type A549 cell lysates with no signal observed at this size in MERTK knockout cell line. To generate this image, wild-type and MERTK knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    • Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Western blot - Anti-MERTK antibody [Y323] (ab52968)
      All lanes : Anti-MERTK antibody [Y323] (ab52968) at 1/500 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : MERTK knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.


      Lanes 1 - 2: Merged signal (red and green). Green - ab52968 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab52968 was shown to specifically react with MERTK in wild-type HAP1 cells as signal was lost in MERTK knockout cells. Wild-type and MERTK knockout samples were subjected to SDS-PAGE. The membrane was blocked with 0. Ab52968 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Western blot - Anti-MERTK antibody [Y323] (ab52968)

      ab52968 used at a 1/200 dilution, 18 hours at 4°C, 5% BSA in Tween PBS.

      Positive cell line: A172

      Negative cell lines: CV1 and HeLa.

      Secondary used is a goat anti-rabbit polyclonal-HRP at 1/10000 dilution.

      Blocked with 5% milk for 1 hour at RT.

      See Abreview

    • Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Western blot - Anti-MERTK antibody [Y323] (ab52968)
      All lanes : Anti-MERTK antibody [Y323] (ab52968) at 1/2000 dilution (purified)

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
      Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG at 1/1000 dilution

      Observed band size: 140-180 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-MERTK antibody [Y323] (ab52968)
      Immunoprecipitation - Anti-MERTK antibody [Y323] (ab52968)

      Lane 1: ab52968 (purified) at 1/20 immunoprecipitating MERTK in HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate.

      Lane 2 - PBS.

      For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling MERTK with purified ab52968 at 1/500.

      Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.

      Negative control using PBS instead of primary antibody (Inset).

    • Immunohistochemistry (Frozen sections) - Anti-MERTK antibody [Y323] (ab52968)
      Immunohistochemistry (Frozen sections) - Anti-MERTK antibody [Y323] (ab52968)

      IHC image of MERTK staining in a section of frozen normal human spleen performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab52968, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Western blot - Anti-MERTK antibody [Y323] (ab52968)
      Anti-MERTK antibody [Y323] (ab52968) at 1/1000 dilution (unpurified) + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Observed band size: 180 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] (ab52968)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lymphoma tissue labeling MERTK with unpurified ab52968 at a 1/50 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-MERTK antibody [Y323] (ab52968)
      Anti-MERTK antibody [Y323] (ab52968)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    文献 (61)

    发表研究结果有使用 ab52968？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab52968 被引用在 61 文献中.

    • Vago JP  et al. Gas6/Axl Axis Activation Dampens the Inflammatory Response in Osteoarthritic Fibroblast-like Synoviocytes and Synovial Explants. Pharmaceuticals (Basel) 16:N/A (2023). PubMed: 37242486
    • Pop OT  et al. AXL Expression on Homeostatic Resident Liver Macrophages Is Reduced in Cirrhosis Following GAS6 Production by Hepatic Stellate Cells. Cell Mol Gastroenterol Hepatol 16:17-37 (2023). PubMed: 37004869
    • Summers RJ  et al. Therapeutic Targeting of MERTK and BCL-2 in T-Cell and Early T-Precursor Acute Lymphoblastic Leukemia. Cancers (Basel) 14:N/A (2022). PubMed: 36551626
    • Kim J  et al. A Bioprinted Bruch's Membrane for Modeling Smoke-Induced Retinal Pigment Epithelium Degeneration via Hybrid Membrane Printing Technology. Adv Healthc Mater 11:e2200728 (2022). PubMed: 35841587
    • Liu Y  et al. N-glycosylation stabilizes MerTK and promotes hepatocellular carcinoma tumor growth. Redox Biol 54:102366 (2022). PubMed: 35728303
    View all Publications for this product

    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

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MERTK antibody [Y323]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Ciliary Body)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6 & TRIS pH 9
    Permeabilization
    Yes - 0.3% Triton X-100
    Specification
    Ciliary Body
    Blocking step
    Serum as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Dylan Windell

    Verified customer

    提交于 May 13 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MERTK antibody [Y323]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Blood vessel-Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH9
    Permeabilization
    No
    Specification
    Blood vessel-Brain
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Nov 30 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MERTK antibody [Y323]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Lung)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
    Permeabilization
    No
    Specification
    Lung
    Blocking step
    PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Nov 14 2017

    Western blot abreview for Anti-MERTK antibody [Y323]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (cell lines)
    Loading amount
    20 µg
    Specification
    cell lines
    Gel Running Conditions
    Reduced Denaturing (4-15% Tris-glycine gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Mr. Nir Drayman

    Verified customer

    提交于 Mar 05 2012

    Question

    Our customer is looking for the isotype control for ab52968 for FACS, ICC/IF and some inhibitory assay. As it was indicated in the reference article of ab52968, also the primary ab ab52968 and its isotype control were purchased from Abcam:



    Carr AJ et al. Molecular characterization and functional analysis of phagocytosis by human embryonic stem cell-derived RPE cells using a novel human retinal assay. Mol Vis 15:283-95 (2009). WB.



    p.285 To examine the role of MERTK in phagocytosis, we incubated HESC-RPE cells with rabbit monoclonal antibody to 1:40 MERTK or 1:40 control rabbit IgG (both Abcam, Cambridge, UK) for 1 h.



    Kindly advise the catalog no. of this isotype control.



    Thanks in advance.

    Read More
    Answer

    Unfortunately, in few cases we cannot find the connection between the antibody used in a publication and the antibody used in our catalog, especially as we are trying to link this kind of information to the datasheets.
    However, as we have only two matching isotypes in this case, and both differ only in the amount we sell:
    https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-ab125938.html https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-ab125938.html.
    https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-pre-diluted-ab128142.html https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-pre-diluted-ab128142.html.

    Read More

    Question

    Phone call requesting the concentration of the Anti-MERTK antibody [Y323] (ab52968)

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    I can now confirm that the concentration of the anti-MERTK antibody [Y323] (ab52968), of the lot number GR54920-5 is 0.1180 mg/ml.

    I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

    Read More

    Question

    Hi, please it is very urgent,
    can you reply?
    Kind regards

    Read More
    Answer

    Thank you for contacting us and sorry for the delay of our reply.
    Please find below the reply I just received from the source of the anti-MERTK antibody [Y323], ab52968:
    > "Sorry for the delay in response. As stated previously, the anti-MERTK antibody ab52968 displayed clear membranous and some cytoplasmic IHC staining of human lymphoma tissue.
    > Yes, it is true despite the region of the immunogen.
    > The image is from formalin-fixed paraffin-embedded tissue, and we performed in-house testing.
    > Although we do not have a paper for IHC, the antibody's specificity is demonstrated in WB and IF.
    > Unfortunately, that is all the information we could find."
    I appreciate the time you have spent on these experiments and would be pleased to arrange, if you agree, a replacement in compensation. The lot we currently have in stock is GR54920. Let me know if you received a different lot so I could send this one to you. Also, could you please inform me when the antibody was ordered and if it was ordered via a distributor?
    Alternatively I can send you another anti-MERTK antibody that could maybe confirm the results you obtained with ab52968. The link to the list of the 9 anti-MERTK we have is: https://www.abcam.com/index.html?t=17227&pt=1
    We would like to investigate this issue much further, so could you please provide some information about your protocol by filling in the attached questionnaire?
    I look forward to your reply and I hope this case will be resolved as quickly as possible.
    Thank you for your cooperation.

    Read More

    Question

    Hi, please it is very urgent,
    can you reply?

    Read More
    Answer


    I just wanted to let you know that we are still waiting for a reply from the originator of this product. I have sent a reminder and I hope we will receive the details about the IHC image you requested in the very near future.
    I am very sorry for the delay. I will forward to you the information as soon as I receive it. Thank you for your patience.

    Read More

    Question

    Esteemed Company,
    I am working with abcam Antibody ab52968 (for Mertk).
    Looking at image reported in your data sheet regarding IHC result, I can see a staining that not appear to be membranous as expected. Can you explain me the result?
    Thank you very much.
    Best regards.

    Read More
    Answer

    Thank you for contacting us. I apologise for the waiting.
    Anti-MERTK antibody [Y323] (ab52968) displayed clear membranous and some cytoplasmic IHC staining of human lymphoma tissue (see larger image attached).
    The Human Protein Atlas states “Most normal tissues showed moderate to strong cytoplasmic positivity with a granular pattern” (see image from the link below).
    http://www.proteinatlas.org/ENSG00000153208
    I also found another example of MER cytoplasmic staining in IHC (attached).
    I hope this information was helpful. Please do not hesitate to contact us back for further assistance.

    Read More

    Question

    The datasheet for this product states that it recognizes both human and mouse however, all of the experimental data provided are performed with human tissues and cells. We need an antibody against mouse MerTK that has been shown to be successful for flow cytometry. If we order this antibody (along with the recomended secondary needed for FC) and we are unsuccessful with mouse samples using flow cytometry, are there any quality assurances? Will you provide supplemental antibody or refund? We are a small laboratory, so we have to stretch our funds. I don't want an investment in this product to be a loss of both valuable time and research funds.

    Read More
    Answer

    Thank you for contacting Abcam. The antibody, ab52968, anti-MERTK antibody [Y323], is covered under our Abpromise, which means that if the antibody does not work as stated on the data sheet and if our support specialist cannot help resolve the issue, then we will provide you with a free of charge replacement or a refund. In this case it is guranteed to work in flow cytometry in mouse tissue. I hope this helps assure about our products and our guaranteed . If there is anything else I can do, please let me know.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com