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使用敲除细胞株进行验证重组RabMAb

重组Anti-Carbonic Anhydrase 9/CA9抗体[EPR4151(2)] (ab108351)

Reviews (4)Q&A (2)References (18)
Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
  • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
  • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
  • Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4151(2)] to Carbonic Anhydrase 9/CA9
  • Suitable for: WB, IHC-P
  • 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

概述

  • 产品名称

    Anti-Carbonic Anhydrase 9/CA9抗体[EPR4151(2)]
    参阅全部 Carbonic Anhydrase 9/CA9 一抗

  • 描述

    兔单克隆抗体[EPR4151(2)] to Carbonic Anhydrase 9/CA9

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IHC-Pmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • IHC: Human stomach tissue; WB: HT-29 cells and human stomach lysate.

  • 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ab108351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 (3)
1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 50 kDa).
IHC-P (1)
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
说明
WB
1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 50 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

靶标

  • 功能

    Reversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia.

  • 组织特异性

    Expressed primarily in carcinoma cells lines. Expression is restricted to very few normal tissues and the most abundant expression is found in the epithelial cells of gastric mucosa.

  • 序列相似性

    Belongs to the alpha-carbonic anhydrase family.
    Contains 1 alpha-carbonic anhydrase domain.

  • 翻译后修饰

    Asn-346 bears high-mannose type glycan structures.

  • 细胞定位

    Nucleus. Nucleus, nucleolus. Cell membrane. Cell projection, microvillus membrane. Found on the surface microvilli and in the nucleus, particularly in nucleolus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • CA-IX antibody
      • CA9 antibody
      • CAH9_HUMAN antibody
      • CAIX antibody
      • Carbonate dehydratase IX antibody
      • Carbonic anhydrase 9 antibody
      • Carbonic anhydrase IX antibody
      • Carbonic dehydratase antibody
      • G250 antibody
      • Membrane antigen MN antibody
      • MN antibody
      • P54/58N antibody
      • pMW1 antibody
      • RCC associated protein G250 antibody
      • RCC-associated antigen G250 antibody
      • Renal cell carcinoma-associated antigen G250 antibody
      see all

    图片

    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      All lanes : Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351) at 1/200 dilution

      Lane 1 : Wild-type HCT 116 cell lysate
      Lane 2 : CA9 knockout HCT 116 cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : HT-29 cell lysate
      Lane 5 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 50 kDa
      Observed band size: 50 kDa



      Western blot: Anti-CA9 antibody [EPR4151(2)] (ab108351) staining at 1/200 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108351 was shown to bind specifically to CA9. A band was observed at 50 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CA9 knockout cell line. To generate this image, wild-type and CA9 knockout HCT 116 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling Carbonic Anhydrase 9/CA9 with Purified ab108351 at 1:150 dilution (0.89 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351) at 1/10000 dilution (Purified) + HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 50 kDa
      Observed band size: 54, 58 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID: 12966427, 30734594 and 15951534).

      Blocking Buffer and concentration: 5% NFDM/TBST

    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351) at 1/10000 dilution (Purified) + Human stomach lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 50 kDa
      Observed band size: 54, 58 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID: 12966427, 30734594 and 15951534).

      Blocking Buffer and concentration: 5% NFDM/TBST

    • Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
      Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)

    文献 (18)

    发表研究结果有使用 ab108351？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab108351 被引用在 18 文献中.

    • Fu Y  et al. Downregulation of HHLA2 inhibits ovarian cancer progression via the NF-κB signaling pathway and suppresses the expression of CA9. Cell Immunol 388-389:104730 (2023). PubMed: 37210768
    • Denize T  et al. Renal cell carcinoma in children and adolescents: a retrospective study of a French-Italian series of 93 cases. Histopathology 80:928-945 (2022). PubMed: 35238063
    • Kallio P  et al. Blocking Angiopoietin-2 Promotes Vascular Damage and Growth Inhibition in Mouse Tumors Treated with Small Doses of Radiation. Cancer Res 80:2639-2650 (2020). PubMed: 32312835
    • Li Y  et al. Dynamic surveillance of tamoxifen-resistance in ER-positive breast cancer by CAIX-targeted ultrasound imaging. Cancer Med 9:2414-2426 (2020). PubMed: 32048471
    • Lorenzo-Pouso AI  et al. Prognostic value of CAIX expression in oral squamous cell carcinoma: a systematic review and meta-analysis. J Enzyme Inhib Med Chem 35:1258-1266 (2020). PubMed: 32466707
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
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    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Carbonic Anhydrase IX antibody [EPR4151(2)]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Gastric cells)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA buffer, pH 9
    Permeabilization
    Yes - Triton 0.025%
    Specification
    Gastric cells
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    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 Jun 03 2015

    Western blot abreview for Anti-Carbonic Anhydrase IX antibody [EPR4151(2)]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Cancer Cell Lines)
    Specification
    Cancer Cell Lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°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.

    Abcam user community

    Verified customer

    提交于 Aug 05 2014

    Western blot abreview for Anti-Carbonic Anhydrase IX antibody [EPR4151(2)]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Sample
    Human Cell lysate - whole cell (Glioma)
    Specification
    Glioma
    Blocking step
    Milk as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 25°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.

    Abcam user community

    Verified customer

    提交于 Dec 27 2013

    Western blot abreview for Anti-Carbonic Anhydrase IX antibody [EPR4151(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30000 cells
    Gel Running Conditions
    Reduced Denaturing (11)
    Sample
    Human Cell lysate - whole cell (MCF-7 cells (human breast adenocarcinoma cell line)
    Specification
    MCF-7 cells (human breast adenocarcinoma cell line
    Treatment
    0.5% oxygen for 16hrs
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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.

    Abcam user community

    Verified customer

    提交于 Nov 05 2013

    Question

    looking for CA-9 mouse antibodies looking for HIF antibodies

    Read More
    Answer

    Thank you for contacting us.

    We have several HIF antibodies in our catalog that react with mouse. For example, ab8366: HIF1 alpha, ab199: HIF2 alpha, ab10134: HIF3 alpha, ab465: HIF1 beta. If you could let me know which HIF you are looking for and what application it would be used for, I would be happy to help you narrow down the results.

    While we do not have any antibodies to Carbonic Anhydrase IX that are known to react with mouse, the antibodies ab108351 and ab15086 are likely to show some cross-reactivity due to sequence homology. If you would like to test either of these antibodies for use in mouse, I would be happy to offer you our 100% testing discount program. If you purchase the antibody for full price and submit an Abreview with your mouse results, you will receive a discount code that can be redeemed for a free primary antibody on a future order.

    I hope this helps, please let me know if you need any additional information or assistance.

    Read More

    Question

    I was wondering if you offer a sample size for your carbonic anhydrase IX monoclonal antibody? I was hoping to test out the antibody prior to purchasing it.

    Read More
    Answer

    Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

    We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

    If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

    Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews

    I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

    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