Anti-RCN1/RCN 抗体 [EPR17163-117] (ab210404)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17163-117] to RCN1/RCN
- Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-RCN1/RCN antibody [EPR17163-117]
RCN1/RCN 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR17163-117] to RCN1/RCN -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- WB: Human placenta and fetal kidney lysates; HEK-293, SH-SY5Y, HeLa, Neuro-2a, C6, PC-12 and NIH/3T3 whole cell lysates; E14 mouse embryo lysate; E14 rat embryo lysate; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates. IHC-P: Human cerebrum and lung adenocarcinoma tissues; Mouse cerebral cortex tissue; Rat hippocampus tissue. ICC/IF: SH-SY5Y, Neuro-2a and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 and SH-SY5Y cells. IP: 293T whole cell lysate.
-
特記事項
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.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR17163-117 -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab210404の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
Flow Cyt (Intra) |
1/700.
|
|
IP |
1/50.
|
|
IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
WB |
1/2000. Detects a band of approximately 44, 46 kDa (predicted molecular weight: 39 kDa).
|
|
ICC/IF |
1/500.
|
特記事項 |
---|
Flow Cyt (Intra)
1/700. |
IP
1/50. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/2000. Detects a band of approximately 44, 46 kDa (predicted molecular weight: 39 kDa). |
ICC/IF
1/500. |
ターゲット情報
-
機能
May regulate calcium-dependent activities in the endoplasmic reticulum lumen or post-ER compartment. -
配列類似性
Belongs to the CREC family.
Contains 6 EF-hand domains. -
細胞内局在
Endoplasmic reticulum lumen. - Information by UniProt
-
参照データベース
- Entrez Gene: 5954 Human
- Entrez Gene: 19672 Mouse
- Entrez Gene: 362182 Rat
- Omim: 602735 Human
- SwissProt: Q15293 Human
- SwissProt: Q05186 Mouse
- Unigene: 97887 Human
- Unigene: 397500 Mouse
see all -
別名
- Epididymis secretory protein Li 84 antibody
- FLJ37041 antibody
- HEL S 84 antibody
see all
画像
-
All lanes : Anti-RCN1/RCN antibody [EPR17163-117] (ab210404) at 1/2000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : RCN1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab210404 observed at 45 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab210404 was shown to react with RCN1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266098 (knockout cell lysate ab258172) was used. Wild-type HEK-293T and RCN1 knockout HEK-293T 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. ab210404 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 2000 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.
-
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on neurons of Human cerebrum is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on SH-SY5Y cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab210404 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
-
All lanes : Anti-RCN1/RCN antibody [EPR17163-117] (ab210404) at 1/5000 dilution
Lane 1 : Human placenta lysate
Lane 2 : Human fetal kidney lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 6 : E14 mouse embryo lysate
Lane 7 : E14 rat embryo lysate
Lane 8 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 39 kDa
Observed band size: 44,46 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 8416973).
-
All lanes : Anti-RCN1/RCN antibody [EPR17163-117] (ab210404) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat heart lysate
Lane 7 : Rat kidney lysate
Lane 8 : Rat spleen lysate
Lane 9 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 10 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 11 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 39 kDa
Observed band size: 44,46 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-8: 1 minute; Lane 9,10 and 11: 10 seconds.
-
Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on tumor cells of Human lung adenocarcinoma is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on neurons of mouse cerebral cortex is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on neurons of rat hippocampus is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on Neuro-2a cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab210404 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on NIH/3T3 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab210404 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RCN1/RCNwith ab210404 at 1/700 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
Cells were permeabilised with 90% methanol (diluted with 1X PBS).
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCNwith ab210404 at 1/700 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
Cells were permeabilised with 90% methanol (diluted with 1X PBS).
-
RCN1/RCN was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab210404 at 1/50 dilution.
Western blot was performed from the immunoprecipitate using ab210404 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: 293T whole cell lysate, 10µg (Input).
Lane 2: ab210404 IP in 293T whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab210404 in 293T whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
Certificate of Compliance
参考文献 (1)
ab210404 は 1 報の論文で使用されています。
- Wang JW et al. RCN1 induces sorafenib resistance and malignancy in hepatocellular carcinoma by activating c-MYC signaling via the IRE1α-XBP1s pathway. Cell Death Discov 7:298 (2021). PubMed: 34663798