AB316306

Anti-CBLL1 antibody [EPR25389-121]

BOND RX™ Validated
20ul selling size
Recombinant
RabMAb
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Rabbit Recombinant Monoclonal HAKAI antibody. Suitable for WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

View Alternative Names

HAKAI, RNF188, CBLL1, E3 ubiquitin-protein ligase Hakai, Casitas B-lineage lymphoma-transforming sequence-like protein 1, RING finger protein 188, RING-type E3 ubiquitin transferase Hakai, c-Cbl-like protein 1

13 Images

Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling CBLL1 with ab316306 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CBLL1 with ab316306 at 1/1000 (0.513 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil (PMID : 19535458). The section was incubated with ab316306 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling CBLL1 with ab316306 at 1/1000 (0.513 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human gastric carcinoma. The section was incubated with ab316306 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling CBLL1 with ab316306 at 1/100 (5.13 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mainly nuclear staining in MCF7 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Supplier Data

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

CBLL1 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab316306 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316306 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab316306 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316306 in MCF7 whole cell lysate

All lanes:

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (ab316306) at 1/30 dilution

All lanes:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling CBLL1 with ab316306 at 1/2000 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung. The section was incubated with ab316306 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CBLL1 with ab316306 at 1/100 (5.13 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in NIH/3T3 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CBLL1 with ab316306 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling CBLL1 with ab316306 at 1/2000 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat liver. The section was incubated with ab316306 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Supplier Data

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

CBLL1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab316306 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316306 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 2 : ab316306 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316306 in NIH/3T3 whole cell lysate

All lanes:

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (ab316306) at 1/30 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 24s

Supplier Data

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CBLL1 antibody [EPR25389-121] (ab316306) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell fresh lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell fresh lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell fresh lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55-60 kDa,36 kDa

false

Exposure time: 37s

Supplier Data

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CBLL1 antibody [EPR25389-121] (ab316306) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting human CBLL1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55-60 kDa,36 kDa

false

Exposure time: 48s

Supplier Data

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CBLL1 antibody [EPR25389-121] (ab316306) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55-60 kDa,36 kDa

false

Exposure time: 92s

Carrier free

Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR25389-121

Isotype

IgG

Carrier free

Reacts with

Human, Mouse, Rat

Applications

WB, Flow Cyt (Intra), ICC/IF, IHC-P, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/1000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/500", "FlowCytIntra-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/2000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/500", "FlowCytIntra-species-notes": "" }, "Rat": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/2000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }

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Product details

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

What are the advantages of a recombinant monoclonal antibody?
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Conditional Ambient

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

CBLL1 also known as Hakai is an E3 ubiquitin ligase. Its molecular weight is approximately 65 kDa and it functions by mediating ubiquitination which leads to the controlled degradation of target proteins. CBLL1 is expressed in several tissues including heart liver and kidney. The protein predominantly functions within the cytoplasm but can also translocate to the nucleus to exert its enzymatic activities.

Biological function summary

CBLL1 plays a significant role in cellular processes such as endocytosis cell adhesion and regulation of epithelial-mesenchymal transition (EMT). CBLL1 functions as part of a complex that helps target proteins for degradation through the ubiquitin-proteasome system. This activity is especially relevant in the regulation of cell signaling pathways where it targets various substrates such as E-cadherin for ubiquitination.

Pathways

CBLL1 is important in modulating signaling cascades like the Wnt and TGF-beta pathways. By influencing these pathways CBLL1 interacts with proteins like beta-catenin and SMADs. The degradation of E-cadherin by CBLL1 facilitates cellular detachment and migration a critical step in cellular processes regulated by these pathways.

CBLL1's involvement in the ubiquitin-proteasome system links it to cancer development and progression. The protein's regulation of EMT and cell adhesion implicates it in cancers such as gastric and colon cancers. Additionally CBLL1 interacts with proteins like beta-catenin in Wnt signaling which is often dysregulated in these cancers. Understanding the function of CBLL1 may offer potential therapeutic targets for combating these malignancies.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

E3 ubiquitin-protein ligase that mediates ubiquitination of several tyrosine-phosphorylated Src substrates, including CDH1, CTTN and DOK1 (By similarity). Targets CDH1 for endocytosis and degradation (By similarity). Associated component of the WMM complex, a complex that mediates N6-methyladenosine (m6A) methylation of RNAs, a modification that plays a role in the efficiency of mRNA splicing and RNA processing (PubMed : 29507755). Its function in the WMM complex is unknown (PubMed : 29507755).

See full target information CBLL1

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

For licensing inquiries, please contact partnerships@abcam.com

Anti-CBLL1 antibody [EPR25389-121]

Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Western blot - Anti-CBLL1 antibody [EPR25389-121] (AB316306)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Primary Antibodies

Primary Antibodies

Cell Lines & Lysates

Cell Lines & Lysates

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise