Recombinant Anti-RAB8A antibody [EPR14873] (ab188574)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14873] to RAB8A
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAB8A antibody [EPR14873]
See all RAB8A primary antibodies -
Description
Rabbit monoclonal [EPR14873] to RAB8A -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, HeLa, 293T, HCT-116, C6, Raw264.7, PC-12 and NIH3T3 lysates and human brain tissue lysate. IHC-P: Human stomach and human adenocarcinoma of colon tissues. ICC/IF: HepG2 cells. Flow Cyt (intra): HeLa cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR14873 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 647 Anti-RAB8A antibody [EPR14873] - C-terminal (ab207548)
- HRP Anti-RAB8A antibody [EPR14873] - C-terminal (ab209046)
- PE Anti-RAB8A antibody [EPR14873] - C-terminal (ab210666)
- Alexa Fluor® 555 Anti-RAB8A antibody [EPR14873] - C-terminal (ab212004)
- Alexa Fluor® 594 Anti-RAB8A antibody [EPR14873] - C-terminal (ab212006)
- Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
- Anti-RAB8A antibody [EPR14873] - BSA and Azide free (ab271954)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab188574 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/260.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (6) |
1/1000 - 1/10000. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
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IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/500.
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Notes |
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Flow Cyt (Intra)
1/260. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa). |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
Target
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Function
May be involved in vesicular trafficking and neurotransmitter release. Together with RAB11A, RAB3IP, the exocyst complex, PARD3, PRKCI, ANXA2, CDC42 and DNMBP promotes transcytosis of PODXL to the apical membrane initiation sites (AMIS), apical surface formation and lumenogenesis. Together with MYO5B and RAB11A participates in epithelial cell polarization. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Cellular localization
Cell membrane. Golgi apparatus. Cytoplasm > perinuclear region. Cell projection. Colocalizes with OPTN at the Golgi complex and in vesicular structures close to the plasma membrane. In the GDP-bound form, present in the perinuclear region. Shows a polarized distribution to distal regions of cell protrusions in the GTP-bound form. Colocalizes with PARD3, PRKCI, EXOC5, OCLN, PODXL and RAB11A in apical membrane initiation sites (AMIS) during the generation of apical surface and luminogenesis. - Information by UniProt
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Database links
- Entrez Gene: 4218 Human
- Entrez Gene: 17274 Mouse
- Entrez Gene: 117103 Rat
- Omim: 165040 Human
- SwissProt: P61006 Human
- SwissProt: P55258 Mouse
- SwissProt: P35280 Rat
- Unigene: 642874 Human
see all -
Alternative names
- AA409338 antibody
- MEL antibody
- Mel transforming oncogene (derived from cell line NK14) RAB8 homolog antibody
see all
Images
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All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RAB8A knockout HeLa cell lysate
Lane 3 : HCT116 cell lysate
Lane 4 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 24 kDaLanes 1-4: Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab188574 Anti-RAB8A antibody [EPR14873] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : RAB8A knockout HAP1 cell lysate
Lane 3 : HCT116 cell lysate
Lane 4 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 24 kDaLanes 1 - 4: Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab188574 was shown to specifically react with RAB8A when RAB8A knockout samples were used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 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 1/10 000 dilution for 1 h at room temperature before imaging. -
Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue labelling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution
Lane 1 : C6 lysate
Lane 2 : RAW264.7 lysate
Lane 3 : PC-12 lysate
Lane 4 : NIH3T3 lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 24 kDa
Observed band size: 24 kDa -
Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution + HCT-116 lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated
Predicted band size: 24 kDa
Observed band size: 24 kDa -
All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/10000 dilution
Lane 1 : HepG2 lysate
Lane 2 : HeLa lysate
Lane 3 : 293T lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 24 kDa
Observed band size: 24 kDa -
Intracellular flow cytometrical analysis of HeLa cells labeling RAB8A with ab188574 at 1/260 (pink) compared to a Isotype control (green). FITC-conjugated goat-anti-rabbit secondary antibody at 1/150 were used for the analysis.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed HepG2 cells labelling RAB8A with ab188574 at 1/500 followed by Goat anti rabbit IgG (Alexa Fluor® 488) and DAPI staining.
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Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (21)
ab188574 has been referenced in 21 publications.
- Nabar NR et al. LRRK2 is required for CD38-mediated NAADP-Ca2+ signaling and the downstream activation of TFEB (transcription factor EB) in immune cells. Autophagy 18:204-222 (2022). PubMed: 34313548
- Zhao CY et al. Acrylamide Exposure Destroys the Distribution and Functions of Organelles in Mouse Oocytes. Front Cell Dev Biol 10:834964 (2022). PubMed: 35295848
- Song T et al. ENKD1 promotes CP110 removal through competing with CEP97 to initiate ciliogenesis. EMBO Rep 23:e54090 (2022). PubMed: 35301795
- Kallemeijn WW et al. Proteome-wide analysis of protein lipidation using chemical probes: in-gel fluorescence visualization, identification and quantification of N-myristoylation, N- and S-acylation, O-cholesterylation, S-farnesylation and S-geranylgeranylation. Nat Protoc 16:5083-5122 (2021). PubMed: 34707257
- Wang H et al. Unraveling GLUT-mediated transcytosis pathway of glycosylated nanodisks. Asian J Pharm Sci 16:120-128 (2021). PubMed: 33613735