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Anti-A2B5抗体[105] (ab53521)

Reviews (2)Q&A (9)References (36)
Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
  • Flow Cytometry (Intracellular) - Anti-A2B5 antibody [105] (ab53521)

Key features and details

  • Mouse monoclonal [105] to A2B5
  • Suitable for: Flow Cyt (Intra), ICC/IF
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgM

概述

  • 产品名称

    Anti-A2B5抗体[105]

  • 描述

    小鼠单克隆抗体[105] to A2B5

  • 宿主

    Mouse

  • 经测试应用

    适用于: Flow Cyt (Intra), ICC/IFmore details

  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Chicken

  • 免疫原

    Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • ICC/IF: Primary mouse neurons/glia, DIV14 cells, Primary rat neurons/glia, DIV14 cells, PC12 cells. Flow Cyt (Intra): SH-SY5Y cells.

  • 常规说明

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab53521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
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab18400 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
ICC/IF (1)
Use a concentration of 1 µg/ml.
说明
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab18400 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
ICC/IF
Use a concentration of 1 µg/ml.

靶标

  • 相关性

    A2B5 is a cell surface ganglioside epitope expressed in developing thymic epithelial cells, oligodendrocyte progenitors and neuroendocrine cells.

  • 细胞定位

    Cell surface

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
    Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)

    ab53521 staining A2B5 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab53521 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150121, Goat polyclonal Secondary Antibody to Mouse IgM - mu chain (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
    Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)

    ab53521 staining A2B5 in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab53521 at 1µg/ml and ab6046. Cells were then incubated with ab150121 at 1/1000 dilution (shown in green) and ab150080 at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
    Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)

    ICC/IF image of ab53521 stained PC12 cells. The cells were 4% formaldehyde fixed (10 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 (ab53521, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgM (H+L) used at a 1/1000 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).

  • Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)
    Immunocytochemistry/ Immunofluorescence - Anti-A2B5 antibody [105] (ab53521)This image is courtesy of an Abreview submitted by Mr Carl Hobbs

    A2B5 antibody [105] - Stem Cell Marker (ab53521) used in immunocytochemical detection on Cor1 (mouse) neuronal stem cells. Cor1 neuronal stem cells were fixed in formaldehyde, permeabilized, blocked in 1% BSA for 30 mins at RT. ab53521 was incubated at 1/500 for 16 hours in TBS/BSA/azide/0.5% Triton. Secondary Antibody:anti mouse IgM conjugated: to Alexa Fluor® 488 (1/1000).

    See Abreview

  • Flow Cytometry (Intracellular) - Anti-A2B5 antibody [105] (ab53521)
    Flow Cytometry (Intracellular) - Anti-A2B5 antibody [105] (ab53521)

    Overlay histogram showing SH-SY5Y cells stained with ab53521 (red line). The cells were fixed with 80% 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 (ab53521, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

文献 (36)

发表研究结果有使用 ab53521？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab53521 被引用在 36 文献中.

  • Zhu Y  et al. Single-Cell Analysis for Glycogen Localization and Metabolism in Cultured Astrocytes. Cell Mol Neurobiol 40:801-812 (2020). PubMed: 31863221
  • Luo X  et al. Fyn gene silencing reduces oligodendrocytes apoptosis through inhibiting ERK1/2 phosphorylation in epilepsy. Artif Cells Nanomed Biotechnol 48:298-304 (2020). PubMed: 31852295
  • Yu Y  et al. Microglial Hv1 proton channels promote white matter injuries after chronic hypoperfusion in mice. J Neurochem 152:350-367 (2020). PubMed: 31769505
  • Matjusaitis M  et al. Reprogramming of Fibroblasts to Oligodendrocyte Progenitor-like Cells Using CRISPR/Cas9-Based Synthetic Transcription Factors. Stem Cell Reports 13:1053-1067 (2019). PubMed: 31708478
  • Naik AA  et al. Intra-generational protein malnutrition impairs temporal astrogenesis in rat brain. Biol Open 6:931-942 (2017). PubMed: 28546341
View all Publications for this product

客户评价及客户问答

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1-10 of 11 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-A2B5 antibody [105] - Stem Cell Marker

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain)
Specification
Brain
Fixative
formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
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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 22 2010

Question

Customer kindly contacted us for assistance in finding the mRNA sequence of this target.

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Answer

I have review the availible literature regarding the A2B5 epitope and have asked the lab to search as well. Unfortunately, we have found no instances recording the mRNA sequence for this target. A2B5 seems to have first been recognized in 1979 in by Eisenbarth et al, Proc. Nat. Acad. Sci. (1979, 76:4913–4917) and has been in use for some time as a marker for gangliosides in developing thymic epithelial cells, oligodendrocyte progenitors and neuroendocrine cells.

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Question

Inquiry: Hi, I'm wondering if the A2B5 (53521)is the same as the ATCC A2B5 clone 105. Thank you.

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Answer

Thank you for your inquiry.

Yes, it does appear these are the same product as they have the same clone number.

I hope this information helps. Please contact us with any other questions.

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Question

No signal from rat insulinoma INS1

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Answer

Thank you for confirming these details and for your cooperation. The details provided 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 ab53521. 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.  

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Question

Thank you so much for your confirmation of our conversation. Following our discussion, I acquired and attempted to label PC-12 cells with the A2B5 antibody and did not have success. Furthermore, I have checked and our secondary antibodies are working. The antibodies we buy are polyclonal and raised again heavy and light chain, meaning that the fact that the A2B5 antibody is IgM should make no difference to the ability of the secondary to target it. What do you need from me in order to replace the vial of A2B5 that I have now?

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Answer

I am sorry this antibody is still not working for you. I will need the Abcam reference number for the order (six digits) or a PO number, or the date of the order and your institution. Did you have an antibody in mind that you would like to replace it with?

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Question

No signal from rat insulinoma INS1

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Answer

Thank you for bringing this issue to our attention. As I mentioned, if the PC12 stain does not work and you have confirmed that the secondary is appropriate (and that it is still fluorescent), I will replace the antibody. There is a chance that a single vial has gone bad. If you also consult the Bartholomeusz 1989 paper I mentioned and which you are already aware of, and any others you think may have an ICC protocol, you may find advice which we do not have on the datasheet.

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Question

I would like to test ab66253 and ab53521 in IHC-P, an untested application.

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Answer

I am very pleased to hear you would like to accept our offer and test ab66253 and ab53521 in IHC-P. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for IHC-P and include this 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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Question

To whom this may concern, I would like to request for Certificates of Compliance for the following antibodies. The product numbers and lot numbers for the antibodies are as follows: Product          Description                                                Lot No. ab53521         Mouse monoclonal [105] to A2B5                  GR8737 ab56069         Mouse monclonal [F16 P2 D8] to...2A6          GR51855 ab62204         Rabbit polyclonal to Cytochrome...2D6           GR6576 Please send these certificates to this email address. I'd be happy to answer any questions or address any concerns regarding this email. Thanks for your time and cooperation.

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Answer

Thank you for contacting Abcam. Attached please find the Certificate of Compliance documents you requested. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Question

My customer plan to use ab53521 in flow cytometer assay. Which product should be chosen as the isotype control for it? Please give us your suggestions. Thank you.

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Answer

Thank you for your patience. The lab has finally isotyped the product and confirmed that it is a kappa isotype. Therefore, the suitable isotype control for this product is: ab18401 Mouse IgM, kappa monoclonal [MM-30] - isotype control Please let me know if there is anything else that I can help you with. Have a nice day!

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Question

I have bought this Ab. I am wondering if I can use anti-mouseIgG secondary or do I have to use anti-mouseIgM secondary Ab? Thank you.

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Answer

Thank you for your enquiry. Since the primary antibody is mouse IgM, you would need to use a secondary antibody that reacts with mouse IgM (anti-Mouse IgM or anti-Mouse IgG (Fab)). I am afraid that a mouse IgG might not suitable. I hope this information helps. If there is anything else that I can help you with, please do not hesitate to contact me.

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1-10 of 11 Abreviews or Q&A

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