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使用敲除细胞株进行验证重组RabMAb

重组Anti-CDC42抗体[EPR15620] (ab187643)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunoprecipitation - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Flow Cytometry (Intracellular) - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
  • Anti-CDC42 antibody [EPR15620] (ab187643)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15620] to CDC42
  • Suitable for: IHC-P, IP, WB, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

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Human CDC42 knockout HEK-293T cell line (ab266522)

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概述

  • 产品名称

    Anti-CDC42抗体[EPR15620]
    参阅全部 CDC42 一抗
  • 描述

    兔单克隆抗体[EPR15620] to CDC42
  • 宿主

    Rabbit
  • 经测试应用

    适用于: IHC-P, IP, WB, Flow Cyt (Intra), ICC/IFmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HEK-293T, HepG2, MCF7, HT-29, Jurkat and HeLa whole cell lysate (ab150035); Mouse brain, Mouse kidney, Mouse spleen and Rat spleen lysates. IP: HT-29 cell lysate. Flow Cyt (intra): HeLa cells. ICC/IF: U937 cells. IHC-P: Human lung carcinoma, breast carcinoma and colon tissues; Rat colon tissue.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EPR15620
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Cytokinesis
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Small G Proteins
    • Other
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Regulation
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Kinases/Phosphatases
    • Other
    • Cancer
    • Signal transduction
    • G protein signaling
    • Small G proteins
    • Ras family

相关产品

  • Alternative Versions

    • Alexa Fluor® 488 Anti-CDC42 antibody [EPR15620] (ab214952)
    • Alexa Fluor® 647 Anti-CDC42 antibody [EPR15620] (ab215234)
    • HRP Anti-CDC42 antibody [EPR15620] (ab215810)
    • Anti-CDC42 antibody [EPR15620] - Low endotoxin, Azide free (ab221781)
    • APC Anti-CDC42 antibody [EPR15620] (ab225345)
    • PE Anti-CDC42 antibody [EPR15620] (ab225346)
    • Anti-CDC42 antibody [EPR15620] - BSA and Azide free (ab271953)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CDC42 knockout HEK-293T cell line (ab266522)
  • KO cell lysates

    • Human CDC42 knockout HEK-293T cell lysate (ab256868)
  • KO cell pellets

    • Human CDC42 knockout HEK-293T cell pellet (ab278988)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • Mouse kidney normal tissue lysate - total protein (ab29305)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Mouse brain tissue lysate - total protein (ab30151)
    • HT-29 whole cell lysate (ab3952)
    • Jurkat whole cell lysate (ab7899)
    • Mouse spleen tissue lysate - total protein (ab7937)
  • Recombinant Protein

    • Recombinant Human CDC42 protein (ab87713)
  • Related Products

    • Recombinant Human CDC42 protein (ab87713)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab187643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
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/20 - 1/40.
WB
1/10000 - 1/50000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
Flow Cyt (Intra)
1/170.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF (1)
1/500.
说明
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/20 - 1/40.
WB
1/10000 - 1/50000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
Flow Cyt (Intra)
1/170.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/500.

靶标

  • 功能

    Plasma membrane-associated small GTPase which cycles between an active GTP-bound and an inactive GDP-bound state. In active state binds to a variety of effector proteins to regulate cellular responses. Involved in epithelial cell polarization processes. Causes the formation of thin, actin-rich surface projections called filopodia.
  • 序列相似性

    Belongs to the small GTPase superfamily. Rho family. CDC42 subfamily.
  • 翻译后修饰

    AMPylation at Tyr-32 and Thr-35 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo.
  • 细胞定位

    Cell membrane.
  • Target information above from: UniProt accession P60953 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: 998 Human
    • Entrez Gene: 12540 Mouse
    • Entrez Gene: 64465 Rat
    • Omim: 116952 Human
    • SwissProt: P60953 Human
    • SwissProt: P60766 Mouse
    • SwissProt: Q8CFN2 Rat
    • Unigene: 467637 Human
    • Unigene: 1022 Mouse
    • Unigene: 447553 Mouse
    • Unigene: 60067 Rat
    see all
  • 形式

    There are 2 isoforms produced by alternative splicing. Isoform 1 also known as: Brain; Isoform 2 also known as: Placental.
  • 别名

    • CDC42 antibody
    • CDC42_HUMAN antibody
    • CDC42Hs antibody
    • Cell division control protein 42 homolog antibody
    • Cell division cycle 42 (GTP binding protein 25kDa) antibody
    • Cell division cycle 42 antibody
    • dJ224A6.1.1 (cell division cycle 42 (GTP-binding protein, 25kD)) antibody
    • dJ224A6.1.2 (cell division cycle 42 (GTP-binding protein, 25kD)) antibody
    • G25K antibody
    • G25K GTP-binding protein antibody
    • Growth regulating protein antibody
    • GTP binding protein 25kDa antibody
    • Small GTP binding protein CDC42 antibody
    • TKS antibody
    see all

图片

  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : CDC42 knockout HEK-293T cell lysate
    Lane 3 : MCF7 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 21 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab187643 Anti-CDC42 antibody [EPR15620] was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody [EPR15620] (ab187643)

    Immunocytochemistry/Immunofluorescence analysis of U937 (Human histiocytic lymphoma cell line) labelling CDC42 with purified ab187643 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : CDC42 knockout HAP1 cell lysate
    Lane 3 : MCF7 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 21 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab187643 was shown to specifically react with CDC42 in wild-type HAP1 cells. No band was observed when CDC42 knockout samples were examined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1/10,000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunoprecipitation - Anti-CDC42 antibody [EPR15620] (ab187643)

    Lane 1 (input): HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate,10μg
    Lane 2(+): HT-29 whole cell lysate
    Lane 3(-): Rabbit monoclonal IgG (ab172730) instead of  ab187643 in HT-29 whole cell lysate

    Ab187643 immunoprecipitating CDC42 in HT-29 whole cell lysate. Capture antibody was used at a 1:60 dilution. For western blotting, ab187643 was used as the primary antibody at a 1:1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST

  • Flow Cytometry (Intracellular) - Anti-CDC42 antibody [EPR15620] (ab187643)
    Flow Cytometry (Intracellular) - Anti-CDC42 antibody [EPR15620] (ab187643)

    Intracellular Flow Cytometry analysis of HeLa cells (human cervix adenocarcinoma epithelial).Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Primary antibody was used at a 1/120 dilution (red). A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (black). Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)

    ab187643 staining CDC42 in Human lung carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections). Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on human lung carcinoma.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)

    ab187643 staining CDC42 in Mouse colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections).  Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on mouse colon.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)

    ab187643 staining CDC42 in Rat colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections).  Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on rat colon.

  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/20000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HT-29 cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

  • Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    Western blot - Anti-CDC42 antibody [EPR15620] (ab187643)
    All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (ab187643)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling CDC42 with ab187643 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

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

  • Anti-CDC42 antibody [EPR15620] (ab187643)
    Anti-CDC42 antibody [EPR15620] (ab187643)

实验方案

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

文献 (33)

发表研究结果有使用 ab187643?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab187643 被引用在 33 文献中.

  • Jiang L  et al. Salmonella Typhimurium reprograms macrophage metabolism via T3SS effector SopE2 to promote intracellular replication and virulence. Nat Commun 12:879 (2021). PubMed: 33563986
  • Zhang A  et al. TNK2 promoted esophageal cancer progression via activating egfr-akt signaling. J Clin Lab Anal 35:e23700 (2021). PubMed: 33484472
  • Engevik AC  et al. Recruitment of Polarity Complexes and Tight Junction Proteins to the Site of Apical Bulk Endocytosis. Cell Mol Gastroenterol Hepatol 12:59-80 (2021). PubMed: 33548596
  • Chen L  et al. MTSS1 inhibits colorectal cancer metastasis by regulating the CXCR4/CXCL12 signaling axis. Int J Mol Med 47:N/A (2021). PubMed: 33649808
  • Yoshida Y  et al. Cdc42 has important roles in postnatal angiogenesis and vasculature formation. Dev Biol 477:64-69 (2021). PubMed: 34019880
View all Publications for this product

客户评价及客户问答

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提交评价 提交问题

Immunocytochemistry/ Immunofluorescence abreview for Anti-CDC42 antibody [EPR15620]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (3T3)
Permeabilization
Yes - triton X100
Specification
3T3
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°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.

Peter Gould

Verified customer

提交于 Nov 08 2016

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

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