Astana Biomed Group, an authorized Abcam distributor in Central Asia

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: ICC, WB, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • 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: ICC, WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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.

  • 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.

    ab255904 is the carrier-free version of ab254147. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab255904 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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
    For more information see here.

Properties

Images

  • Western blot - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Western blot - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    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.

  • Immunocytochemistry - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Immunocytochemistry - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)

    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).

  • Immunoprecipitation - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Immunoprecipitation - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)

    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).

  • Western blot - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Western blot - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    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 1/100000 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.

  • Immunocytochemistry - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Immunocytochemistry - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)

    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).

  • Immunoprecipitation - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Immunoprecipitation - Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)

    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).

  • Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)
    Anti-RHOG antibody [1F3-RHOG] - BSA and Azide free (ab255904)

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