重组Anti-PDCD4抗体[EPR3431] (ab80590)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3431] to PDCD4
- Suitable for: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-PDCD4抗体[EPR3431]
参阅全部 PDCD4 一抗 -
描述
兔单克隆抗体[EPR3431] to PDCD4 -
宿主
Rabbit -
经测试应用
适用于: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human PDCD4 (N terminal). The exact sequence is proprietary.
-
阳性对照
- WB: HEK293, Jurkat, HEK293 and HeLa cell lysates. IHC-P: human breast cancer tissue, human colon tissue; ICC/IF: HeLa cells. IP: HeLa cells. Flow Cyt (intra): HeLa cells.
-
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 3.10 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR3431 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab80590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 |
---|---|---|
WB |
1/1000. Predicted molecular weight: 52 kDa.
For unpurified use: 1/20000.00000 - 1/100000.00000 |
|
IP |
1/20.
For unpurified use: 1/30.00000 |
|
IHC-P |
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
|
ICC/IF |
1/50.
For unpurified use: 1/100.00000 - 1/250.00000 |
说明 |
---|
WB
1/1000. Predicted molecular weight: 52 kDa. For unpurified use: 1/20000.00000 - 1/100000.00000 |
IP
1/20. For unpurified use: 1/30.00000 |
IHC-P
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
ICC/IF
1/50. For unpurified use: 1/100.00000 - 1/250.00000 |
靶标
-
功能
Inhibits translation initiation and cap-dependent translation. May excert its function by hindering the interaction between EIF4A1 and EIF4G. Inhibits the helicase activity of EIF4A. Modulates the activation of JUN kinase. Down-regulates the expression of MAP4K1, thus inhibiting events important in driving invasion, namely, MAPK85 activation and consequent JUN-dependent transcription. May play a role in apoptosis. Tumor suppressor. Inhibits tumor promoter-induced neoplastic transformation. Binds RNA. -
组织特异性
Up-regulated in proliferative cells. Highly expressed in epithelial cells of the mammary gland. Reduced expression in lung cancer and colon carcinoma. -
序列相似性
Belongs to the PDCD4 family.
Contains 2 MI domains. -
结构域
Binds EIF4A1 via both MI domains. -
翻译后修饰
Polyubiquitinated, leading to its proteasomal degradation. Rapidly degraded in response to mitogens. Phosphorylation of the phosphodegron promotes interaction with BTRC and proteasomal degradation. -
细胞定位
Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm. Predominantly nuclear under normal growth conditions, and when phosphorylated at Ser-457. Exported from the nucleus in the absence of serum. - Information by UniProt
-
数据库链接
- Entrez Gene: 27250 Human
- Omim: 608610 Human
- SwissProt: Q53EL6 Human
- Unigene: 711490 Human
-
别名
- Death up-regulated gene protein antibody
- Dug antibody
- H731 antibody
see all
图片
-
All lanes : Anti-PDCD4 antibody [EPR3431] (ab80590) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa -
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PDCD4 with Purified ab80590 at 1:50 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling PDCD4 with Purified ab80590 at 1:3000 dilution (0.05 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling PDCD4 with Purified ab80590 at 1:3000 dilution (0.05 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
-
All lanes : Anti-PDCD4 antibody [EPR3431] (ab80590) at 1/20000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PDCD4 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab80590 observed at 51 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab80590 was shown to react with PDCD4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261833 (knockout cell lysate ab257278) was used. Wild-type HeLa and PDCD4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab80590 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 20000 dilution and a 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.
-
All lanes : Anti-PDCD4 antibody [EPR3431] (ab80590) at 1/20000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PDCD4 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 52 kDaLanes 1 - 4: Merged signal (red and green). Green - ab80590 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab80590 was shown to specifically react with PDCD4 in wild-type HAP1 cells as signal was lost in PDCD4 knockout cells. Wild-type and PDCD4 knockout samples were subjected to SDS-PAGE. ab80590 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C both at 1/20000 dilution. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
-
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PDCD4 with purified ab80590 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10µg
Lane 2 (+): ab80590 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab80590 in HeLa whole cell lysate
Ab80590 (Purified) at 1/30 dilution (20 µg/ml) immunoprecipitating PDCD4 in HeLa whole cell lysate. For western blotting, ab80590 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (22)
ab80590 被引用在 22 文献中.
- Huang G et al. Gender Differential Expression of AR/miR-21 Signaling Axis and Its Protective Effect on Renal Ischemia-Reperfusion Injury. Front Cell Dev Biol 10:861327 (2022). PubMed: 35573679
- Gong X et al. A smart multiantenna gene theranostic system based on the programmed assembly of hypoxia-related siRNAs. Nat Commun 12:3953 (2021). PubMed: 34172725
- Huang WC et al. The MEK/ERK/miR-21 Signaling Is Critical in Osimertinib Resistance in EGFR-Mutant Non-Small Cell Lung Cancer Cells. Cancers (Basel) 13:N/A (2021). PubMed: 34885115
- Sun J et al. microRNA-93, upregulated in serum of nasopharyngeal carcinoma patients, promotes tumor cell proliferation by targeting PDCD4. Exp Ther Med 19:2579-2587 (2020). PubMed: 32256737
- Guo T et al. miR-590-5p may regulate colorectal cancer cell viability and migration by targeting PDCD4. Exp Ther Med 20:55 (2020). PubMed: 32952645