| 產(chǎn)品編號 | bs-5216R |
| 英文名稱 | [KO驗證抗體] Phospho-Bad (Ser118) Rabbit pAb |
| 中文名稱 | 磷酸化相關死亡促進因子抗體 |
| 別 名 | Bad(phospho S118); Bad(phospho Ser118); p-Bad(S118); p-Bad(Ser118); p-Bad(phospho Ser118); BBC 2; BBC2; BBC6; Bcl 2 Antagonist of Cell Death; Bcl 2 Binding Component 6; BCL X/BCL 2 Binding Protein; BCL X Binding Protein; Bcl XL/Bcl 2 Associated Death Prom |
| 產(chǎn)品類型 | 磷酸化抗體 KO驗證抗體 |
| 研究領域 | 腫瘤 細胞生物 免疫學 神經(jīng)生物學 信號轉導 細胞凋亡 線粒體 |
| 抗體來源 | Rabbit |
| 克隆類型 | Polyclonal |
| 交叉反應 | Human,Mouse,Rat (predicted: Rabbit,Pig,Cow,Dog,Horse) |
| 產(chǎn)品應用 | WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=2ug/Test
not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
| 理論分子量 | 22 kDa |
| 檢測分子量 | |
| 細胞定位 | 細胞漿 細胞膜 線粒體 |
| 性 狀 | Liquid |
| 濃 度 | 1mg/ml |
| 免 疫 原 | KLH conjugated Synthesised phosphopeptide derived from human BAD around the phosphorylation site of Ser118: RM(p-S)DE |
| 亞 型 | IgG |
| 純化方法 | affinity purified by Protein G |
| 緩 沖 液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 保存條件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事項 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| PubMed | PubMed |
| 產(chǎn)品介紹 |
Bad is a member of the Bcl2 family and acts to promote apoptosis by forming heterodimers with the survival proteins Bcl2 and BclxL, thus preventing them from binding with BAX. Bad is found on the outer mitochondrial membrane and, once phosphorylated in response to growth stimuli, translocates to the cytoplasm. The phosphorylation status of Bad represents a key checkpoint for death or cell survival. JNK-induced phosphorylation of BAD serine 128 promotes the apoptotic role of Bad by opposing the inhibitory effect of growth factor on Bad-mediated apoptosis. Cdc2-induced phosphorylation of Bad serine 128 has an inhibitory effect on its interaction with 14-3-3 proteins. The latter interaction is critical for Bad phosphorylation at serine 155, a site within the BH3 domain that leads to the release of BclxL and the promotion of cell survival. Alternative splicing of this gene results in two transcript variants which encode the same isoform. Function: Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2. Appears to act as a link between growth factor receptor signaling and the apoptotic pathways. Subunit: Forms heterodimers with the anti-apoptotic proteins, Bcl-X(L), Bcl-2 and Bcl-W. Also binds protein S100A10. The Ser-75/Ser-99 phosphorylated form binds 14-3-3 proteins. Interacts with AKT1 and PIM3. Subcellular Location: Mitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm. Tissue Specificity: Expressed in a wide variety of tissues. Post-translational modifications: Phosphorylated on one or more of Ser-75, Ser-99, Ser-118 andSer-134 in response to survival stimuli, which blocks itspro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75promotes heterodimerization with 14-3-3 proteins. This interactionthen facilitates the phosphorylation at Ser-118, a site within theBH3 motif, leading to the release of Bcl-X(L) and the promotion ofcell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation. Phosphorylation at Ser-99 by PKB/AKT1 is almost completely blockedby the apoptotic C-terminus cleavage product of PKN2 generated bycaspases-3 activity during apoptosis. Methylation at Arg-94 and Arg-96 by PRMT1 inhibits Akt-mediated phosphorylation at Ser-99. Similarity: Belongs to the Bcl-2 family. SWISS: Q92934 Gene ID: 572 Database links: Entrez Gene: 572 Human Entrez Gene: 12015 Mouse Omim: 603167 Human SwissProt: Q92934 Human SwissProt: Q61337 Mouse Unigene: 370254 Human Unigene: 4387 Mouse Unigene: 36696 Rat BAD是BCL2/BAX、BCL-XL/BAX異二聚體的負調節(jié)基因。BAD是BCL2/BCL-XL相關死亡促進因子，作為BCL2、 bCL-XL異二聚體伴分子而促進細胞凋亡。 |
| 產(chǎn)品圖片 |
Sample:
Hela(Human) Cell Lysate at 30 ug
Hela KO Bad (Human) Cell Lysate at 30 ug
Primary: Anti- Phospho-Bad (Ser118) (bs-5216R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-Bad (Ser118) (bs-5216R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD
Sample:
Raw264.7(Mouse) Cell Lysate at 30 ug
Primary: Anti-Phospho-Bad (Ser118) ? (bs-5216R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD
Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control (Black line): Jurkat (Black).
Primary Antibody (green line): Rabbit Anti-Bad(Ser118) antibody (bs-5216R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 15 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-Phospho-Bad (Ser118) antibody (bs-5216R)
Dilution: 2ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature.Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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| 1、抗體溶解方法 | |
| 2、抗體修復方式 | |
| 3、常用試劑的配制 | |
| 4、免疫組化操作步驟 | |
| 5、免疫組化問題解答 | |
| 6、Western Blotting 操作步驟 | |
| 7、Western Blotting 問題解答 | |
| 8、關于肽鏈的設計 | |
| 9、多肽的溶解與保存 | |
| 10、酶標抗體效價測定程序 | |
