來源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實際分子量 |
Rabbit | WB, IF | H, M, R, Mk | 18/23kDa | 25kDa |
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檢測或其它適當(dāng)用途時的一抗稀釋。
建議抗體使用時的稀釋比例如下(實際使用時需根據(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 | - | 1:50-1:200 | - | - | - | - | - |
抗體詳細(xì)信息如下::
About this Antibody | |
Name | ARHGDIA/RhoGDI Rabbit Polyclonal Antibody (KO Validated) |
Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
Isotype | IgG |
Purification method | Affinity purification |
Positive samples | HeLa, COS7, PC-12, Mouse lung, Mouse spleen, Mouse kidney |
Cellular location | Cytoplasm |
Customer validation | WB (Mouse) |
About the Immunogen | |
Immunogen | Recombinant fusion protein of human ARHGDIA (NP_001172006.1). |
Sequence | MAEQEPTAEQLAQIAAENEEDEHSVNYKPPAQKSIQEIQELDKDDESLRKYKEALLGRVAVSADPNVPNVVVTGLTLVCSSAPGPLELDLTGDLESFKKQSFVLKEGVEYRIKISFRVNREIVSGMKYIQHTYRKGVKIDKTDYMVGSYGPRAEEYEFLTPVEEAPKGMLARGSYSIKSRFTDDDKTDHLSWEWNLTIKKDWKD |
Gene ID | 396 |
Swiss Prot | P52565 |
Synonyms | ARHGDIA; GDIA1; HEL-S-47e; NPHS8; RHOGDI; RHOGDI-1; rho GDP-dissociation inhibitor 1 |
Category | MAPK/Erk in GPCR Signaling; SAPK/JNK Signaling; TGF-β Signaling; Jak/Stat:IL-6 Signaling |
Background | Rho family small GTPases, including Rho, Rac and cdc42, act as molecular switches, regulating processes such as cell migration, adhesion, proliferation and differentiation. They are activated by guanine nucleotide exchange factors (GEFs), which catalyze the exchange of bound GDP for GTP, and inhibited by GTPase activating proteins (GAPs), which catalyze the hydrolysis of GTP to GDP. A third level of regulation is provided by the stoichiometric binding of Rho GDP dissociation inhibitor (RhoGDI). RhoGDI affects Rho activity by inhibiting nucleotide exchange and membrane association, regulating activity and localization. The inhibitory and shuttling functions of RhoGDI have been uncoupled using mutant forms of RhoGDI. Phosphorylation of GDIs and/or GTPases can modulate their affinity for each other and, therefore, GTPase mediated signaling. PAK1 phosphorylation of RhoGDI at serines 101 and 174 causes release and activation of Rac1, but not RhoA. |
包裝清單:
產(chǎn)品編號 | 產(chǎn)品名稱 | 包裝 |
AF5117 | ARHGDIA/RhoGDI Rabbit Polyclonal Antibody (KO Validated) | 50μl |
AZ050 | Western一抗稀釋液 | 50ml |
— | 說明書 | 1份 |
保存條件:
ARHGDIA/RhoGDI 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,長期不使用可以考慮-20ºC保存,但凍融可能會導(dǎo)致出現(xiàn)輕微的渾濁和少量不溶物。
注意事項:
如果本抗體用于Western blot (WB)、免疫熒光(IF)、免疫細(xì)胞化學(xué)(ICC)等實驗,請注意回收使用過的稀釋抗體。回收的抗體通常至少可以重復(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。如果回收的抗體出現(xiàn)明顯的絮狀物或長霉長菌等情況,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效果,請考慮使用上述檢測專用的一抗稀釋液。
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,請穿實驗服并戴一次性手套操作。
使用說明:
請根據(jù)抗體的實際用途選擇相應(yīng)的使用方法。
1. Western檢測:
a. 按照推薦的稀釋比例用Western一抗稀釋液稀釋抗體。
b. 把經(jīng)過封閉的蛋白膜與稀釋好的一抗4℃緩慢搖動過夜或室溫緩慢搖動2小時,確保稀釋的抗體至少能在搖動的瞬間覆蓋蛋白膜。
c. 回收稀釋的一抗,4℃保存以備下次繼續(xù)使用。
d. 按照Western的實驗步驟進(jìn)行后續(xù)的洗滌、二抗孵育、洗滌和檢測等操作。
2. 免疫染色:
可以使用免疫染色一抗稀釋液(P0103)稀釋抗體,使用后注意回收稀釋好的一抗,
3. 其它實驗操作請自行參考適當(dāng)?shù)膒rotocol進(jìn)行。
4. 代表性圖片:
Fig. 1. Western blot analysis of extracts of various cell lines, using ARHGDIA 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). | Fig. 2. Western blot analysis of extracts from normal (control) and ARHGDIA knockout (KO) 293T cells, using ARHGDIA 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. Immunofluorescence analysis of HeLa cells using ARHGDIA antibody. Blue: DAPI for nuclear staining. |