Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)

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

Be the first to review this product! Submit a review

Images

Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (AB271185), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/30	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 Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info 1/500	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information Clathrin light chain A

Function

Clathrin is the major protein of the polyhedral coat of coated pits and vesicles. Acts as a component of the TACC3/ch-TOG/clathrin complex proposed to contribute to stabilization of kinetochore fibers of the mitotic spindle by acting as inter-microtubule bridge (PubMed:15858577, PubMed:21297582).

Alternative names

Clathrin light chain A, Lca, CLTA

Recommended products

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR24231-72
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
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

Notes

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.

Supplementary info

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

Activity summary

Clathrin light chain also known as clathrin LCa and LCb is a low-molecular-weight component of the clathrin protein complex. It plays a mechanical role in the processes that drive clathrin-mediated endocytosis. Each clathrin triskelion the basic unit of the clathrin coat comprises three clathrin heavy chains and three clathrin light chains. The mass of clathrin light chain is approximately 25-29 kDa. This protein is expressed in various tissues throughout the body with notable presence in the central nervous system and liver.

Biological function summary

Clathrin light chain modulates the lattice dynamics by interacting with clathrin heavy chains in a clathrin protein complex. This interaction supports the stability and assembly of the clathrin lattice affecting the efficiency and specificity of endocytic vesicle formation. Clathrin light chain acts in concert with other accessory proteins to regulate clathrin coat assembly and disassembly playing a dynamic role in cellular trafficking processes.

Pathways

The clathrin light chain participates in essential cellular pathways such as the clathrin-mediated endocytosis and synaptic vesicle recycling pathways. These pathways facilitate the internalization and recycling of membrane proteins and lipids ensuring cellular homeostasis. In these processes the clathrin light chain works closely with related proteins such as dynamin and adaptin to control vesicle scission and cargo selection.

Associated diseases and disorders

Abnormalities in clathrin light chain function connect to neurodegenerative disorders like Huntington's disease and certain types of cancers. In Huntington's disease disrupted endocytic pathways that involve clathrin light chain impact cellular communication and nutrient uptake. Cancer cells may exhibit altered expression or function of clathrin light chain affecting pathways important for cell proliferation and survival. The clathrin light chain can interact with the huntingtin protein in these contexts influencing disease progression and cellular dynamics.

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.

13 product images

Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
False colour image of Western blot: Anti-Clathrin light chain antibody [EPR24231-72] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab271185 was shown to bind specifically to Clathrin light chain. A band was observed at 35 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in CLTA knockout cell line Human CLTA (Clathrin light chain A) knockout HEK-293T cell line ab267334 (knockout cell lysate Human CLTA (Clathrin light chain A) knockout HEK-293T cell lysate ab258366). To generate this image, wild-type and CLTA knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: CLTA knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human CLTA (Clathrin light chain A) knockout HEK-293T cell line (Human CLTA (Clathrin light chain A) knockout HEK-293T cell line ab267334)
Lane 3: A431 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 27 kDa
Observed band size: 35 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Clathrin light chain with ab271185 at 1/5000 (0.102 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in human cerebrum. The section was incubated with ab271185 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Clathrin light chain was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab271185 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271185 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: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab271185 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab271185 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
All lanes: Immunoprecipitation - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Predicted band size: 27 kDa
Observed band size: 35 kDa
Flow Cytometry (Intracellular) - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma cell) cells labelling Clathrin light chain with ab271185 at 1/500 dilution (0.1ug)(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). A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF cells labelling Clathrin light chain with ab271185 at 1/250 (2.036 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MEF cells is observed. 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 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID: 31672988, 24852344).
Exposure time: 70 seconds.
All lanes: Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg
Lane 5: C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg
Lane 6: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Lane 7: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 27 kDa
Observed band size: 35 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Clathrin light chain with ab271185 at 1/5000 (0.102 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in mouse cerebrum. The section was incubated with ab271185 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Clathrin light chain with ab271185 at 1/250 (2.036 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells is observed. 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 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This antibody does not recognize human CLTB.
Exposure time: 70 seconds.
All lanes: Western blot - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185) at 1/1000 dilution
Lane 1: Human Clathrin light chain recombinant protein at 0.01 µg
Lane 2: Human CLTB recombinant protein at 0.01 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 27 kDa
Observed band size: 35 kDa
Immunoprecipitation - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Clathrin light chain was immunoprecipitated from 0.35 mg MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate with ab271185 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271185 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: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate 10 ug
Lane 2: ab271185 IP in MEF whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab271185 in MEF whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
All lanes: Immunoprecipitation - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Predicted band size: 27 kDa
Observed band size: 35 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling Clathrin light chain with ab271185 at 1/5000 (0.102 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in rat cerebrum. The section was incubated with ab271185 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 cells labelling Clathrin light chain with ab271185 at 1/250 (2.036 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in C2C12 cells is observed. 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 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Clathrin light chain antibody [EPR24231-72] (ab271185)
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling Clathrin light chain with ab271185 at 1/500 dilution (0.1ug)(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). A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.