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Rabbit Recombinant Monoclonal RBX1 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Expected
Tested
Mouse
Predicted
Predicted
Predicted
Expected
Predicted
Rat
Predicted
Predicted
Predicted
Expected
Predicted

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Predicted
Predicted

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Predicted
Predicted

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Predicted
Predicted

Species

Mouse, Rat

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Predicted
Predicted

Species

Mouse, Rat

Dilution info

-

Notes

-

Target data

Function

E3 ubiquitin ligase component of multiple cullin-RING-based E3 ubiquitin-protein ligase (CRLs) complexes which mediate the ubiquitination and subsequent proteasomal degradation of target proteins, including proteins involved in cell cycle progression, signal transduction, transcription and transcription-coupled nucleotide excision repair (PubMed:10230407, PubMed:10579999, PubMed:15983046, PubMed:16678110, PubMed:19112177, PubMed:19679664, PubMed:23455478, PubMed:27565346, PubMed:29769719, PubMed:11961546, PubMed:22748924). CRLs complexes and ARIH1 collaborate in tandem to mediate ubiquitination of target proteins, ARIH1 mediating addition of the first ubiquitin on CRLs targets (PubMed:27565346). The functional specificity of the E3 ubiquitin-protein ligase complexes depends on the variable substrate recognition components. As a component of the CSA complex promotes the ubiquitination of ERCC6 resulting in proteasomal degradation. Recruits the E2 ubiquitin-conjugating enzyme CDC34 to the complex and brings it into close proximity to the substrate. Probably also stimulates CDC34 autoubiquitination. May be required for histone H3 and histone H4 ubiquitination in response to ultraviolet and for subsequent DNA repair. Promotes the neddylation of CUL1, CUL2, CUL4 and CUL4 via its interaction with UBE2M. Involved in the ubiquitination of KEAP1, ENC1 and KLHL41. In concert with ATF2 and CUL3, promotes degradation of KAT5 thereby attenuating its ability to acetylate and activate ATM.

Alternative names

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Rabbit Recombinant Monoclonal RBX1 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR6850(B)

Purification technique

Affinity purification Protein A

Dissociation constant

2.18 x 10-11 M

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab248556 is the carrier-free version of ab133565.

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

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.

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.

Supplementary info

Biological function summary

RBX1 associates with the cullin-RING ubiquitin ligase (CRL) complex. It interacts with cullin proteins and other regulatory subunits to promote the polyubiquitination and subsequent degradation of target proteins. This regulatory activity influences many cellular processes including cell cycle progression DNA repair and signal transduction. By ensuring the degradation of proteins at the right time RBX1 maintains cellular homeostasis.

Activity summary

RBX1 also known as RING-box protein 1 or ROC1 acts as an E3 ubiquitin ligase part of the ubiquitin-proteasome system. This protein facilitates the transfer of ubiquitin from E2 enzymes to target substrates marking them for degradation. The molecular weight of RBX1 is approximately 12 kDa. RBX1 is mainly expressed in various human tissues with significant expression in the heart brain and skeletal muscle. Its expression in these tissues suggests a wide involvement in cellular processes.

Pathways

RBX1 plays an important role in the control of protein stability through the ubiquitin-proteasome pathway. This pathway is important for regulating key cellular functions like the entrainment of the cell cycle and apoptosis. RBX1 also interacts with proteins like SKP1 and CUL1 as part of the SCF complex which targets specific proteins for degradation. By modulating these pathways RBX1 contributes to the overall regulation of cellular growth and stress responses.

Associated diseases and disorders

RBX1's malfunctioning links to various types of cancer and neurodegenerative diseases. In cancer altered RBX1 activity can lead to the inappropriate degradation of tumor suppressor proteins promoting uncontrolled cell division. RBX1 is also implicated in neurodegenerative disorders where misregulation of protein degradation contributes to the accumulation of damaged proteins exacerbating disease progression. Additionally the interaction of RBX1 with proteins like p53 in cancer and tau proteins in Alzheimer's illustrates its potential role in disease pathogenesis.

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.

7 product images

  • Immunoprecipitation - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Immunoprecipitation - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.

    Purified ab133565 at 1:20 dilution (1μg) immunoprecipitating RBX1 in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg.

    Lane 2 (+): ab133565 + Jurkat whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133565 in Jurkat whole cell lysate.

    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1:1000 dilution) was used for Western blotting.

    Blocking Buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM/TBST.

    Observed band size: 13 kDa

    All lanes: Immunoprecipitation - Anti-RBX1 antibody [EPR6850(B)] (AB133565)

    Predicted band size: 12 kDa

  • Western blot - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Western blot - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Image produced using the purified version.

    All lanes: Western blot - Anti-RBX1 antibody [EPR6850(B)] (AB133565) at 1/5000 dilution

    Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

    Lane 2: Mouse heart lysate at 15 µg

    Lane 3: Rat heart lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 12 kDa

    Observed band size: 13 kDa

  • Flow Cytometry (Intracellular) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling RBX1 with Purified ab133565 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RBX1 with Purified ab133565 at 1:50 dilution (3.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat breast tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse breast tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR6850(B)] - BSA and Azide free (ab248556)

    This data was developed using ab133565, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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