Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)

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

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Images

Western blot - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (AB316307), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IP	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Tested	Tested
Rat	Tested	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Target data

See full target information CBLL1

Function

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).

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

Recommended products

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR25389-121
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab316307 is the carrirer-free version of Anti-CBLL1 antibody [EPR25389-121] ab316306.

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

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

Activity summary

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.

Associated diseases and disorders

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.

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:
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13 product images

Western blot - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-CBLL1 antibody [EPR25389-121] (Anti-CBLL1 antibody [EPR25389-121] ab316306) at 1/1000 dilution
Lane 1: Mouse liver tissue lysate at 20 µg with 5% NFDM/TBST
Lane 2: Mouse lung tissue lysate at 20 µg with 5% NFDM/TBST
Lane 3: Rat liver tissue lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 55-60 kDa, 36 kDa
Exposure time: 92s
Western blot - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-CBLL1 antibody [EPR25389-121] (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 with 5% NFDM/TBST
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell fresh lysate at 20 µg with 5% NFDM/TBST
Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell fresh lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 55-60 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-CBLL1 antibody [EPR25389-121] (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 with 5% NFDM/TBST
Lane 2: HeLa transfected with siRNA specifically targeting human CBLL1 whole cell lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 55-60 kDa, 36 kDa
Exposure time: 48s
Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/100 (5.13 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/100 (5.13 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] 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 Anti-CBLL1 antibody [EPR25389-121] 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] 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 Anti-CBLL1 antibody [EPR25389-121] 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] 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 Anti-CBLL1 antibody [EPR25389-121] 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling CBLL1 with Anti-CBLL1 antibody [EPR25389-121] 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 Anti-CBLL1 antibody [EPR25389-121] 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
Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
CBLL1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lane 2: Anti-CBLL1 antibody [EPR25389-121] ab316306 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CBLL1 antibody [EPR25389-121] ab316306 in NIH/3T3 whole cell lysate
All lanes: Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (Anti-CBLL1 antibody [EPR25389-121] ab316306) at 1/30 dilution
All lanes: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with 5% NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 24s
Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] - BSA and Azide free (ab316307)
This data was developed using Anti-CBLL1 antibody [EPR25389-121] ab316306, the same antibody clone in a different buffer formulation.
CBLL1 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CBLL1 antibody [EPR25389-121] ab316306 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

Lane 2: Anti-CBLL1 antibody [EPR25389-121] ab316306 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CBLL1 antibody [EPR25389-121] ab316306 in MCF7 whole cell lysate
All lanes: Immunoprecipitation - Anti-CBLL1 antibody [EPR25389-121] (Anti-CBLL1 antibody [EPR25389-121] ab316306) at 1/30 dilution
All lanes: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s