Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3043] to Rab4 - Early Endosome Marker
- Suitable for: WB, IP, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
产品名称Anti-Rab4抗体[EPR3043] - Early Endosome Marker
参阅全部 Rab4 一抗
描述兔单克隆抗体[EPR3043] to Rab4 - Early Endosome Marker
经测试应用适用于: WB, IP, Flow Cyt, ICC/IFmore details
种属反应性与反应: Mouse, Rat, Human
Synthetic peptide corresponding to Human Rab4 aa 150-250 (C terminal).
- WB: MCF7, PC12, Neuro 2a, 293T, SH SY5Y and Human fetal brain lysates; ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: MCF7 cells.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
存放说明Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), PBS, 0.05% BSA
Concentration information loading...
纯度Protein A purified
Our Abpromise guarantee covers the use of ab109009 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 24 kDa.|
|IP||1/10 - 1/100.|
|ICC/IF||1/170 - 1/1000.|
功能Protein transport. Probably involved in vesicular traffic.
序列相似性Belongs to the small GTPase superfamily. Rab family.
翻译后修饰Phosphorylated by CDK1 kinase during mitosis.
细胞定位Membrane. Cytoplasm. Generally associated with membranes. Cytoplasmic when phosphorylated by CDK1.
- Information by UniProt
- HRES 1 / RAB4 antibody
- Oncogene RAB4 antibody
- Rab 4 antibody
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Rab4 with Purified ab109009 at 1:170 dilution (10 µg/ml). Cells were fixed in 100% Methanol. 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 nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Rab4 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human fetal brain lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109009 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109009 was shown to specifically react with Rab4 when Rab4 knockout samples were used. Wild-type and Rab4 knockout samples were subjected to SDS-PAGE. ab109009 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 2 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lysates/proteins at 1/15 dilution per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 24 kDa
ab109009 (purified) at 1:80 dilution (2µg) immunoprecipitating Rab4 in MCF7 whole cell lysate.
Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab109009 & MCF7 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109009 in MCF7 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Rab4 with Purified ab109009 at 1:200 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. 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).
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling Rab4 with purified ab109009 at 1/250. Cells were fixed with 100% methanol and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
All lanes : Anti-Rab4 antibody [EPR3043] - Early Endosome Marker (ab109009) at 1/1000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : PC12 cell lysate
Lane 3 : Neuro 2a cell lysate
Lane 4 : 293T cell lysate
Lane 5 : SH SY5Y cell lysate
Lane 6 : Human fetal brain lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 24 kDa
ab109009 at 1/500 dilution staining Rab4 in HeLa by Immunofluorescence.
ab109009 被引用在 6 文献中.
- Rivero-Ríos P et al. Distinct Roles for RAB10 and RAB29 in Pathogenic LRRK2-Mediated Endolysosomal Trafficking Alterations. Cells 9:N/A (2020). PubMed: 32709066
- Gong B et al. Sec14l3 potentiates VEGFR2 signaling to regulate zebrafish vasculogenesis. Nat Commun 10:1606 (2019). PubMed: 30962435
- Rivero-Ríos P et al. The G2019S variant of leucine-rich repeat kinase 2 (LRRK2) alters endolysosomal trafficking by impairing the function of the GTPase RAB8A. J Biol Chem 294:4738-4758 (2019). PubMed: 30709905
- Johnson IRD et al. A Paradigm in Immunochemistry, Revealed by Monoclonal Antibodies to Spatially Distinct Epitopes on Syntenin-1. Int J Mol Sci 20:N/A (2019). PubMed: 31795513
- Xin X et al. Drug-delivering-drug platform-mediated potent protein therapeutics via a non-endo-lysosomal route. Theranostics 8:3474-3489 (2018). PubMed: 30026860
- Niu Y et al. Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory. Elife 6:N/A (2017). PubMed: 28134614