Recombinant Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [1F3-RHOG] to RHOG - BSA and Azide free
- Suitable for: WB, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free
See all RHOG primary antibodies -
Description
Mouse monoclonal [1F3-RHOG] to RHOG - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: WB, IP, 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: Jurkat, MDA-MB-231, K562, J774A.1 and PC-12 whole cell lysate. Mouse bone marrow tissue lysate. Rat thymus tissue lysate. ICC/IF: Jurkat and J774A.1 cells. IP: Jurkat and J774A.1 whole cell lysates.
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General notes
ab255904 is the carrier-free version of ab254147.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
1F3-RHOG -
Isotype
IgG2b -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255904 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 |
---|---|---|
WB |
Use at an assay dependent concentration. Detects a band of approximately 18 kDa (predicted molecular weight: 21 kDa).
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 18 kDa (predicted molecular weight: 21 kDa). |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Required for the formation of membrane ruffles during macropinocytosis. Required for the formation of cup-like structures during trans-endothelial migration of leukocytes. In case of Salmonella enterica infection, activated by SopB and ARHGEF26/SGEF, which induces cytoskeleton rearrangements and promotes bacterial entry. -
Sequence similarities
Belongs to the small GTPase superfamily. Rho family. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 391 Human
- Entrez Gene: 56212 Mouse
- Entrez Gene: 308875 Rat
- Omim: 179505 Human
- SwissProt: P84095 Human
- SwissProt: P84096 Mouse
- Unigene: 501728 Human
- Unigene: 259795 Mouse
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Alternative names
- ARHG antibody
- Ras homolog family member G antibody
- Rho G antibody
see all
Images
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All lanes : Anti-RHOG antibody [1F3-RHOG] (ab254147) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : RHOG knockout HEK-293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-RHOG antibody [1F3-RHOG] staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab254147 was shown to bind specifically to RHOG. A band was observed at 18 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in RHOG knockout cell line ab266753 (knockout cell lysate ab263328). To generate this image, wild-type and RHOG knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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All lanes : Anti-RHOG antibody [1F3-RHOG] (ab254147) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 4 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 21 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression molecular weight observed is consistent with what has been described in the literature (PMID:18505794).
Exposure time: 3 minutes.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat cells labelling RHOG with ab254147 at 1/50 dilution, followed by ab150113 goat anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Jurkat cells. ab179513 anti-beta Tubulin rabbit monoclonal antibody at a 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/500 dilution was used to counterstain tubulin (Red). The nuclear counterstain was DAPI (Blue).
Negative control 1: ab254147 at a 1/50 dilution followed by ab150080 at a 1/500 dilution.
Negative control 2: ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
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RHOG was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte), whole cell lysate with ab254147 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254147 at 1/1000 dilution. Mouse IgG for IP (HRP) (ab131368) was used at 1/1000 dilution.
Lane 1: Jurkat whole cell lysate 10µg.
Lane 2: ab254147 IP in Jurkat whole cell lysate.
Lane 3: Mouse monoclonal IgG2b instead of ab254147 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
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All lanes : Anti-RHOG antibody [1F3-RHOG] (ab254147) at 1/1000 dilution
Lane 1 : Mouse bone marrow tissue lysate
Lane 2 : Rat thymus tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 21 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized J774A.1 cells labelling RHOG with ab254147 at 1/50 dilution, followed by ab150113 goat anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in J774A.1 cells. ab179513 anti-beta Tubulin rabbit monoclonal antibody at a 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/500 dilution was used to counterstain tubulin (Red). The nuclear counterstain was DAPI (Blue).
Negative control 1: ab254147 at a 1/50 dilution followed by ab150080 at a 1/500 dilution.
Negative control 2: ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
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RHOG was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate with abab254147 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254147 at 1/1000 dilution. Mouse IgG for IP (HRP) (ab131368) was used at 1/1000 dilution.
Lane 1: J774A.1 whole cell lysate 10µg.
Lane 2: ab254147 IP in J774A.1 whole cell lysate.
Lane 3: Mouse monoclonal IgG2b instead of ab254147 in J774A.1 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254147).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (1)
ab255904 has been referenced in 1 publication.
- Meller J et al. Endogenous RhoG is dispensable for integrin-mediated cell spreading but contributes to Rac-independent migration. J Cell Sci 121:1981-9 (2008). PubMed: 18505794