Anti-RHOG antibody [1F3-RHOG] (ab254147)
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
- Suitable for: WB, IP, ICC
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-RHOG antibody [1F3-RHOG]
See all RHOG primary antibodies -
Description
Mouse monoclonal [1F3-RHOG] to RHOG -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species ICC MouseHumanIP MouseHumanWB MouseRatHuman -
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: Jurkat and J774A.1 cells. IP: Jurkat and J774A.1 whole cell lysates.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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 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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
1F3-RHOG -
Isotype
IgG2b -
Research areas
Images
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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?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.
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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.
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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?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.
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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.
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