Anti-Calponin 3 antibody [EPR28430-76] - BSA and Azide free
- BOND RX™ Validated
- RabMAb
- Recombinant
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
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
- 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.
- 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.
- 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.
- 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
- 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.
- 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.
- 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.
- 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
- 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
- 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
- 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
- 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
- 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
Related conjugates and formulations (1)
-
Anti-Calponin 3 antibody [EPR28430-76]
Reactivity data
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
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Product promise
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