Anti-RAC1 + Cdc42 (phospho S71)抗体
参阅全部 RAC1 + Cdc42 一抗
兔多克隆抗体to RAC1 + Cdc42 (phospho S71)
Rabbit
Cdc 42 [pS71] (100% homologous) and Rho A/B/C [pS73] (92% homologous) are expected to react.
适用于: IHC-P, WBmore details
与反应: Human
Synthetic peptide corresponding to Human RAC1 + Cdc42 (phospho S71). The sequence is conserved in human and mouse RAC 1, 2, and 3, and Cdc 42 human, mouse, rat, dog and frog.
WB: A431 cells treated with EGF. IHC-P: human skin, human stomach tissues
RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA
BSA is IgG and protease free
浓度
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Immunogen affinity purified
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated RAC 1. The final product is generated by affinity chromatography using a RAC 1 derived peptide that is phosphorylated at serine 71.
RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity.
多克隆
IgG
Abpromise™承诺保证使用ab5482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 | 1/20 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
WB | (1) | 1/1000. Detects a band of approximately 23 kDa. |
Entrez Gene: 5879 Human
Entrez Gene: 998 Human
Omim: 116952 Human
Omim: 602048 Human
SwissProt: P60953 Human
SwissProt: P63000 Human
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Western blot - Anti-RAC1 + Cdc42 (phospho S71) antibody (ab5482)
Peptide Competition and Phosphatase Treatment:
Lysates prepared from A431 cells left unstimulated (1) or stimulated with EGF (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either not treated (1-5) or treated with lambda phosphatase (6), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5482 antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2, 6), the nonphosphopeptide corresponding to the immunogen (3), a generic phosphoserine containing peptide (4), or, the phosphopeptide immunogen (5), After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the peptide corresponding to ab5482 blocks the antibody signal. The data also shows that phosphatase stripping eliminates the signal, verifying that the antibody is pho
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAC1 + Cdc42 (phospho S71) antibody (ab5482)
Immunohistochemiscal analysis of paraffin-embedded human skin tissue labeling RAC1 + Cdc42 (phospho S71) with ab5482 at 1/100 dilution (right) compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab5482 diluted in 3% BSA-PBS at a dilution of 1/100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAC1 + Cdc42 (phospho S71) antibody (ab5482)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling RAC1 + Cdc42 (phospho S71) with ab5482 at 1/20 dilution (right) compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab5482 diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.