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AB314330

Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free

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Rabbit Recombinant Monoclonal Calponin 3 antibody. Carrier free. Suitable for WB, IHC-P, IP, ICC/IF and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples.

View Alternative Names

Calponin-3, CNN3

13 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Calponin 3 with ab314329 at 1/4000 (0.128 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on chorionic villi of human placenta (PMID : 20861310). The section was incubated with ab314329 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Calponin 3 with ab314329 at 1/100 (5.1 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 cytoplasmic staining in HeLa cell line. Negative control : MCF7 (PMID : 30911294).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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling Calponin 3 with ab314329 at 1/4000 (0.128 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human skeletal muscle. The section was incubated with ab314329 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IP

Supplier Data

Immunoprecipitation - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Calponin 3 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab314329 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314329 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab314329 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314329 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/30 dilution

All lanes:

HeLa (human cervical 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: 24s

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, 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 Calponin 3 with ab314329 at 1/100 (5.1 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 cytoplasmic staining in NIH/3T3 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Calponin 3 with ab314329 at 1/4000 (0.128 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse colon. The section was incubated with ab314329 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Calponin 3 with ab314329 at 1/4000 (0.128 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat colon. The section was incubated with ab314329 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • IP

Supplier Data

Immunoprecipitation - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Calponin 3 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab314329 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314329 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 : ab314329 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314329 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) 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: 3s

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • WB

Supplier Data

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates in lane 2-5 were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands around 25 kDa are unknown.

All lanes:

Western blot - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat testis tissue lysate at 20 µg

Secondary

Lanes 1 - 5:

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 1/2000 dilution

Lanes 1 - 5:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Observed band size: 40 kDa

false

Exposure time: 15s

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • WB

Supplier Data

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In lanes 5-7, lysates were stored at -80℃ prior to Western Blotting. In lanes 1-4, lysate were freshly made and used for Western Blotting immediately to minimize protein degradation, showing a reduction in the intensity of bands between 15 kDa and 30 kDa.

All lanes:

Western blot - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/1000 dilution

Lanes 1 and 6:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 5:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 7:

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

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) 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: 40 kDa

false

Exposure time: 10s

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • WB

Supplier Data

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : MCF7(PMID : 30911294). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands around 25 kDa are unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) 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: 40 kDa

false

Exposure time: 15s

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • WB

Supplier Data

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : Mouse skeletal muscle Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands around 25 kDa are unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle 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: 40 kDa

false

Exposure time: 26s

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)
  • WB

Supplier Data

Western blot - Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free (AB314330)

This data was developed using ab314329, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The recombinant proteins of Calponin 1, Calponin 2 and Calponin 3 were made in house. This antibody does not react with human Calponin 1 and Calponin 2. In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Calponin 3 antibody [EPR28430-76] (<a href='/en-us/products/primary-antibodies/calponin-3-antibody-epr28430-76-ab314329'>ab314329</a>) at 1/1000 dilution

Lane 1:

His-tagged human Calponin 2 recombinant protein at 20 ng

Lane 2:

His-tagged human Calponin 1 recombinant protein at 20 ng

Lane 3:

His-tagged human Calponin 3 recombinant fragment at 20 ng

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: 17 kDa

false

Exposure time: 15s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR28430-76

Isotype

IgG

Carrier free

Yes

Reacts with

Human, Mouse, Rat

Applications

IP, IHC-P, ICC/IF, WB

applications

Immunogen

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

Specificity

This antibody does not react with human Calponin 1 and Calponin 2.

Reactivity data

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Product details

ab314330 is the carrier-free version of ab314329.

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Supplementary information

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

Calponin 3 also known as CNN3 is a protein that regulates actin cytoskeleton dynamics. It is a smaller protein with a mass of approximately 37.5 kDa. Calponin 3 is expressed in various tissues including smooth muscle and certain non-muscle cells. This protein functions by interacting with actin filaments where it plays a role in stabilizing the cytoskeletal structure. CNN3 may also have similar functions to those of other members of the calponin family engaging in processes that influence cellular contractility and motility.
Biological function summary

The functions of calponin 3 extend to cellular processes such as migration and proliferation. It does not work alone but is a part of a network of proteins and may form complexes with other cytoskeletal components. Calponin 3 contributes to cellular activities by modulating actin filament interactions helping maintain cell shape and structural integrity under stress. As part of the calponin family its function resembles that of other family members which bind with actin and affect the contractile apparatus in cells.

Pathways

Calponin 3 integrates into the regulation of the actin cytoskeleton and cell migration pathways. It is associated with the Rho GTPase pathway influencing actin dynamics through these interactions. Another related pathway includes the calcium signaling pathway as calponin family members often show calcium-sensitive binding to actin. In these pathways calponin 3 interacts with proteins like tropomyosin and myosin which are critical to cytoskeletal organization and movement.

Calponin 3 has relevance to certain cancers and cardiovascular disorders. Changes in calponin 3 expression levels or function have been linked to tumor progression as its role in cell motility might influence cancer metastasis. The protein might also impact cardiovascular disease through its involvement in smooth muscle contraction regulation which could affect blood vessel function and contribute to hypertension. Interaction with myosin light chain kinase can further explain these effects as disruptions in this relationship can exacerbate disease states.

Product protocols

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

Target data

Thin filament-associated protein that is implicated in the regulation and modulation of smooth muscle contraction. It is capable of binding to actin, calmodulin and tropomyosin. The interaction of calponin with actin inhibits the actomyosin Mg-ATPase activity.
See full target information CNN3

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

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