來源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實(shí)際分子量 |
Rabbit | WB, IF, IHC | H, M, R | 43kDa | 53kDa |
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk, Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀釋液,可以用于Western檢測(cè)或其它適當(dāng)用途時(shí)的一抗稀釋。
建議抗體使用時(shí)的稀釋比例如下(實(shí)際使用時(shí)需根據(jù)抗原水平的高低作適當(dāng)調(diào)整):
WB | IP | IF | IHC | ICC | FC | ChIP | ChIP-seq |
1:500-1:2000 | - | 1:50-1:100 | 1:50-1:200 | 1:50-1:200 | - | - | - |
抗體詳細(xì)信息如下::
About this Antibody | |
Name | p53 Rabbit Polyclonal Antibody (KO Validated) |
Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
Isotype | IgG |
Purification method | Affinity purification |
Positive samples | A-431, MCF7, Mouse thymus |
Cellular location | Cytoplasm, Endoplasmic reticulum, Mitochondrion matrix, Nucleus, PML body |
Customer validation | - |
About the Immunogen | |
Immunogen | A synthetic synthetic peptide of mouse p53 (NP_035770.2). |
Sequence | SAKRALPTCTSASPPQKKKPLDGEYFTLKIRGRKRFEMFRELNEALELKDAHATEESGDSRAHSSYLKTKKGQSTSRHKKTMVKKVGPDSD |
Gene ID | 22059 |
Swiss Prot | P02340 |
Synonyms | TP53; P53; TRP53; tumor protein p53; bbl; bfy; bhy; p44 |
Category | p53; Regulation of Apoptosis; PI3K/Akt Signaling; p38 MAPK Signaling |
Background | The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis. p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro. DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to a reduced interaction between p53 and its negative regulator, the oncoprotein MDM2. MDM2 inhibits p53 accumulation by targeting it for ubiquitination and proteasomal degradation. p53 can be phosphorylated by ATM, ATR, and DNA-PK at Ser15 and Ser37. Phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and activation of p53 in response to DNA damage. Chk2 and Chk1 can phosphorylate p53 at Ser20, enhancing its tetramerization, stability, and activity. p53 is phosphorylated at Ser392 in vivo and by CAK in vitro. Phosphorylation of p53 at Ser392 is increased in human tumors and has been reported to influence the growth suppressor function, DNA binding, and transcriptional activation of p53. p53 is phosphorylated at Ser6 and Ser9 by CK1δ and CK1ε both in vitro and in vivo. Phosphorylation of p53 at Ser46 regulates the ability of p53 to induce apoptosis. Acetylation of p53 is mediated by p300 and CBP acetyltransferases. Inhibition of deacetylation suppressing MDM2 from recruiting HDAC1 complex by p19 (ARF) stabilizes p53. Acetylation appears to play a positive role in the accumulation of p53 protein in stress response. Following DNA damage, human p53 becomes acetylated at Lys382 (Lys379 in mouse) in vivo to enhance p53-DNA binding. Deacetylation of p53 occurs through interaction with the SIRT1 protein, a deacetylase that may be involved in cellular aging and the DNA damage response. |
包裝清單:
產(chǎn)品編號(hào) | 產(chǎn)品名稱 | 包裝 |
AF5258 | p53 Rabbit Polyclonal Antibody (KO Validated) | 50μl |
AZ050 | Western一抗稀釋液 | 50ml |
— | 說明書 | 1份 |
保存條件:
p53 Rabbit Polyclonal Antibody (KO Validated) -20ºC保存,Western一抗稀釋液-20ºC或4ºC保存,一年有效。Western一抗稀釋液優(yōu)先推薦4ºC保存,長(zhǎng)期不使用可以考慮-20ºC保存,但凍融可能會(huì)導(dǎo)致出現(xiàn)輕微的渾濁和少量不溶物。
注意事項(xiàng):
如果本抗體用于Western blot (WB)、免疫熒光(IF)、免疫細(xì)胞化學(xué)(ICC)等實(shí)驗(yàn),請(qǐng)注意回收使用過的稀釋抗體;厥盏目贵w通常至少可以重復(fù)使用5-10次。稀釋后的抗體,包括已經(jīng)使用過的稀釋抗體,請(qǐng)4℃保存。
回收后重復(fù)使用的抗體,使用方法同新鮮稀釋的抗體。如果在重復(fù)使用過程中發(fā)現(xiàn)抗體出現(xiàn)輕微混濁現(xiàn)象,可以10,000g離心1-3分鐘,取上清用于后續(xù)檢測(cè)。如果回收的抗體出現(xiàn)明顯的絮狀物或長(zhǎng)霉長(zhǎng)菌等情況,則可以考慮廢棄該抗體。
提供的Western一抗稀釋液也可以用于免疫熒光(IF)、免疫組化(IHC)、免疫細(xì)胞化學(xué)(ICC)等適當(dāng)用途。如果希望獲得最佳的檢測(cè)效果,請(qǐng)考慮使用上述檢測(cè)專用的一抗稀釋液。
本產(chǎn)品僅限于專業(yè)人員的科學(xué)研究用,不得用于臨床診斷或治療,不得用于食品或藥品,不得存放于普通住宅內(nèi)。
為了您的安全和健康,請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。
4. 代表性圖片:
Fig. 1. Western blot analysis of extracts of various cell lines, using p53 antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s. | Fig. 2. Western blot analysis of extracts from normal (control) and p53 knockout (KO) 293T cells, using p53 antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s. |
Fig. 3. Immunohistochemistry of paraffin-embedded human lung cancer using p53 antibody at dilution of 1:100 (40x lens). | Fig. 4. Immunofluorescence analysis of A549 cells using p53 antibody. |