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使用敲除细胞株进行验证重组RabMAb

重组Anti-RhoGDI抗体[EPR3772] (ab108977)

Submit a review Q&A (3)References (2)
Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
  • Anti-RhoGDI antibody [EPR3772] (ab108977)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3772] to RhoGDI
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-RhoGDI抗体[EPR3772]
    参阅全部 RhoGDI 一抗

  • 描述

    兔单克隆抗体[EPR3772] to RhoGDI

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WBmore details
    不适用于: Flow Cyt,IHC-P or IP

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Rat

  • 免疫原

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

  • 阳性对照

    • WB: HEK293T, Jurkat, HeLa, HL-60 and NIH/3T3 cell lysates.

  • 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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab108977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
1/1000 - 1/5000. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).
说明
WB
1/1000 - 1/5000. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).
应用说明
Is unsuitable for Flow Cyt,IHC-P or IP.

靶标

  • 功能

    Regulates the GDP/GTP exchange reaction of the Rho proteins by inhibiting the dissociation of GDP from them, and the subsequent binding of GTP to them.

  • 序列相似性

    Belongs to the Rho GDI family.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • ARHGDIA antibody
    • fa96g11 antibody
    • GDIA 1 antibody
    • GDIA1 antibody
    • GDIR1_HUMAN antibody
    • MGC117248 antibody
    • NPHS8 antibody
    • Rho GDI 1 antibody
    • Rho GDI alpha antibody
    • Rho GDI antibody
    • Rho GDP dissociation inhibitor (GDI) alpha antibody
    • Rho GDP dissociation inhibitor 1 antibody
    • Rho GDP dissociation inhibitor alpha antibody
    • Rho GDP-dissociation inhibitor 1 antibody
    • Rho-GDI alpha antibody
    • RhoGDI 1 antibody
    • RhoGDI alpha antibody
    • RHOGDI antibody
    • RhoGDI1 antibody
    • wu:fa96g11 antibody
    • zgc:55554 antibody
    • zgc:77681 antibody
    see all

图片

  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    All lanes : Anti-RhoGDI antibody [EPR3773] (ab133248) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : ARHGDIA knockout HEK293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 23 kDa



    ab108977 was shown to react with aRHGDIA in wild-type HEK293T cells in Western blot with loss of signal observed in ARHGDIA knockout cell line ab266447 (ARHGDIA knockout cell lysate ab257356). Wild-type HEK293T and ARHGDIA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab108977 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.

    These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    All lanes : Anti-RhoGDI antibody [EPR3772] (ab108977) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : ARHGDIA knockout HEK293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 23 kDa
    Observed band size: 23 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab108977 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab108977 Anti-RhoGDI antibody [EPR3772] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab108977 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    All lanes : Anti-RhoGDI antibody [EPR3772] (ab108977) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : ARHGDIA knockout HEK293T cell lysate
    Lane 3 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 23 kDa
    Observed band size: 27 kDa why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab108977 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab108977 Anti-RhoGDI antibody [EPR3772] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266447 (knockout cell lysate ab257356) was used.  Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE.  ab108977 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    Western blot - Anti-RhoGDI antibody [EPR3772] (ab108977)
    All lanes : Anti-RhoGDI antibody [EPR3772] (ab108977) at 1/1000 dilution

    Lane 1 : Jurkat cell lysates
    Lane 2 : HeLa cell lysates
    Lane 3 : HL-60 cell lysates
    Lane 4 : NIH/3T3 cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 23 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?

  • Anti-RhoGDI antibody [EPR3772] (ab108977)
    Anti-RhoGDI antibody [EPR3772] (ab108977)

文献 (2)

发表研究结果有使用 ab108977？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab108977 被引用在 2 文献中.

  • Kempf SJ  et al. The cognitive defects of neonatally irradiated mice are accompanied by changed synaptic plasticity, adult neurogenesis and neuroinflammation. Mol Neurodegener 9:57 (2014). WB . PubMed: 25515237
  • Yentrapalli R  et al. The PI3K/Akt/mTOR pathway is implicated in the premature senescence of primary human endothelial cells exposed to chronic radiation. PLoS One 8:e70024 (2013). Human . PubMed: 23936371

客户评价及客户问答

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1-3 of 3 Abreviews or Q&A

Question

The only purchase order number i can find is FMVS1125183, but i'm not sure
if this is an internal university number, or if it relates to Abcam's
system as well.
As of today's date (18th Sept 2012), the antibody is no longer in the
6-month guarantee period because of the protracted nature of this process,
however the drop in antibody quality did occur within the 6-month period.
Cheers,

Read More
Answer

Thank you for your email.

Unfortunately the order is out of Abpromise guarantee period. The order was placed on 17th January 2012, which means it was under 6 month guarantee period until 16th June however the complaint was submitted in August so I am sorry we will not be able to replace the product on this occasion. I would suggest contacting us within 6 month period for claiming the guarantee period.

We now have new lot of this product in stock. Please buy the new lot and use it as per your convenience, if in any case the product fails please let us know we will be happy to replace the product.

I hope this information is nevertheless helpful to you. Should you have any question please do not hesitate to contact me.

Read More

Question

I stored it in its original white-topped Abcam vial in my -20 degree
freezer along with my other antibody stocks. There was a slight drop in
quality with the antibody after about 4 months (using 1:5000 dilution), but
from about 5 months onwards it was a sudden drop off to no signal at all,
even at 1:1000.
Cheers,

Read More
Answer

Thanks you for your email. I apologize for the delay.

We unfortunately are unable to find reason why the activity of this antibody is dropping. Also we haven't received any other quality complaint for this antibody so I am sorry we will not be retesting this product at the moment.

To proceed further in this case; could you please provide the purchase order number? I assume that the product you have purchase is within 6 month guarantee period and might be eligible for replacement. If the product was purchased within 6 month guarantee period I will be happy replacing the product.

I will look forward to receiving the order number soon.

Read More

Question

I apologise for the delay in responding. Here is my completed questionnaire.
Cheers,

Read More
Answer

Thank you very much for sending the information.

In order to understand the problem better could you provide details how this antibody was stored?

Was the antibody degradation gradual or it just stopped working after 6 months?

Many thanks for your cooperation in advance. I will look forward to hearing from you soon.

Read More

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