Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)

Rabbit Recombinant Monoclonal RAB8A antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

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Images

Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (AB243568), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

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Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information RAB8A

Function

The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different sets of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion. That Rab is involved in polarized 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 (PubMed:20890297). Regulates the compacted morphology of the Golgi (PubMed:26209634). Together with MYO5B and RAB11A participates in epithelial cell polarization (PubMed:21282656). Also involved in membrane trafficking to the cilium and ciliogenesis (PubMed:21844891, PubMed:30398148). Together with MICALL2, may also regulate adherens junction assembly (By similarity). May play a role in insulin-induced transport to the plasma membrane of the glucose transporter GLUT4 and therefore play a role in glucose homeostasis (By similarity). Involved in autophagy (PubMed:27103069). Participates in the export of a subset of neosynthesized proteins through a Rab8-Rab10-Rab11-dependent endososomal export route (PubMed:32344433). Targeted to and stabilized on stressed lysosomes through LRRK2 phosphorylation (PubMed:30209220). Suppresses stress-induced lysosomal enlargement through EHBP1 and EHNP1L1 effector proteins (PubMed:30209220).

Alternative names

MEL, RAB8, RAB8A, Ras-related protein Rab-8A, Oncogene c-mel

Recommended products

Rabbit Recombinant Monoclonal RAB8A antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: MJF-R22-79-3
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab243568 is the carrier-free version of Anti-RAB8A antibody [MJF-R22-79-3] ab241061.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Collaborations
This antibody was developed with support from The Michael J. Fox Foundation.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RAB8A is a small GTPase also referred to as Ras-related protein Rab-8A involved in intracellular membrane trafficking. It plays a critical role in the regulation of exocytosis the process by which cells release substances to the extracellular space. RAB8A mainly functions by cycling between an inactive GDP-bound form and an active GTP-bound form facilitating vesicle budding transport and fusion with target membranes. This protein is expressed in various tissues including the brain and pancreas where it assists in important cellular functions. RAB8A has a molecular mass of approximately 24 kDa.

Biological function summary

The small GTPase RAB8A contributes to the biosynthetic transport from the trans-Golgi network to the plasma membrane. It performs this activity in coordination with other proteins as part of a larger complex. RAB8A also plays an essential role in cilia formation and maintenance affecting cell polarity and signaling functions. The protein's ability to regulate vesicle movement makes it important for cellular homeostasis and communication.

Pathways

The regulatory function of RAB8A is significant in the ciliary membrane trafficking pathway and the insulin signaling pathway. Within these pathways RAB8A interacts closely with other proteins like Rab11 and Rabin8 which help support its roles in directing vesicle transport. These interactions are essential for proper cellular signaling and the maintenance of polarized cellular structures further supporting the function of various cellular pathways.

Associated diseases and disorders

The dysfunction of RAB8A has associations with certain ciliopathies notably Bardet-Biedl syndrome. This condition involves abnormalities in cilia structure and function leading to symptoms like kidney disease vision impairment and obesity. Moreover disruptions in RAB8A have connections with insulin resistance implicating it in metabolic disorders. In the context of these diseases proteins such as BBS proteins and Rab11 play a role in mediating the effects of dysfunctional RAB8A-related pathways.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
This data was developed using the same antibody clone in a different buffer formulation (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Lanes 1-4: Merged signal (red and green). Green - Anti-RAB8A antibody [MJF-R22-79-3] ab241061 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-RAB8A antibody [MJF-R22-79-3] ab241061 Anti-RAB8A antibody [MJF-R22-79-3] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human RAB8A knockout HeLa cell line ab264993 (knockout cell lysate Human RAB8A knockout HeLa cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab Anti-RAB8A antibody [MJF-R22-79-3] ab241061 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-RAB8A antibody [MJF-R22-79-3] (Anti-RAB8A antibody [MJF-R22-79-3] ab241061) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: RAB8A knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human RAB8A knockout HeLa cell line (Human RAB8A knockout HeLa cell line ab264993)
Lane 3: HCT116 cell lysate at 20 µg
Lane 4: Human brain tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 24 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Immunohistochemical analysis of paraffin-embedded human breast tissue labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human breast (PMID: 14581456). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
RAB8A was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
Lane 1: A549 whole cell lysate 10 μg (Input).
Lane 2: Anti-RAB8A antibody [MJF-R22-79-3] ab241061 IP in A549 whole cell lysate .
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RAB8A antibody [MJF-R22-79-3] ab241061 in A549 whole cell lysate .
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
All lanes: Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] (Anti-RAB8A antibody [MJF-R22-79-3] ab241061)
Predicted band size: 24 kDa
Flow Cytometry (Intracellular) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cell line labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human bladder cancer (PMID: 14581456). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in A549 cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Immunofluorescent analysis of 3% paraformaldehyde-fixed, 0.1% Saponin permeabilized A549 (human lung carcinoma cell line) and RAB8A knock-out A549 cells labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/100 dilution.
This image is kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).
Flow Cytometry (Intracellular) - Anti-RAB8A antibody [MJF-R22-79-3] - BSA and Azide free (ab243568)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cell line labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB8A antibody [MJF-R22-79-3] ab241061).