Antibodies, Proteins, Kits and Reagents for Life Science

150 768 ₸ Product size

100 µl 150 768 ₸

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Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [MJF-R22-79-3] to RAB8A
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
Knockout validated
Reacts with: Human

Product name

Anti-RAB8A antibody [MJF-R22-79-3]
See all RAB8A primary antibodies
Description

Rabbit monoclonal [MJF-R22-79-3] to RAB8A
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant full length protein within Human RAB8A aa 1 to the C-terminus. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.
Database link: P61006

Run BLAST with Run BLAST with
Positive control
- WB: HEK-293T, HCT116 and HeLa whole cell lysates. IHC-P: Human bladder cancer and breast tissues. ICC/IF: A549 and HeLa cells. Flow Cyt: A549 cells. IP: A549 whole cell lysate.
General notes
This antibody was developed with support from The Michael J. Fox Foundation.

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

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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

MJF-R22-79-3
Isotype

IgG
Research areas

Western blot - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

All lanes : Anti-RAB8A antibody [MJF-R22-79-3] (ab241061) 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 kDa

Lanes 1-4: Merged signal (red and green). Green - ab241061 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

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 ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab ab241061 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.
Western blot - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

All lanes : Anti-RAB8A antibody [MJF-R22-79-3] (ab241061) 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 kDa

Lanes 1-4: Merged signal (red and green). Green - ab241061 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

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 ab264994 (knockout cell lysate ab257196) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab241061 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.
Western blot - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

All lanes : Anti-RAB8A antibody [MJF-R22-79-3] (ab241061) at 1/1000 dilution

Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 24 kDa

Exposure time : 7.75 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 15673612).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling RAB8A with 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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling RAB8A with 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.

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] (ab241061)

RAB8A was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with ab241061 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab241061 at 1/1000 dilution. 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: ab241061 IP in A549 whole cell lysate .

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab241061 in A549 whole cell lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 8 seconds.
Flow Cytometry - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cell line labeling RAB8A with ab241061 at 1/600 dilution (red) compared with a 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) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RAB8A with ab241061 at 1/500 dilution followed by 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) (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) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB8A with ab241061 at 1/500 dilution followed by 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) (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) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

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 ab241061 at 1/100 dilution.

This image is kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.
Anti-RAB8A antibody [MJF-R22-79-3] (ab241061)

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