Overview

  • Product name
    Anti-CACNA1C antibody [L57/46]
    See all CACNA1C primary antibodies
  • Description
    Mouse monoclonal [L57/46] to CACNA1C
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IHC-Fr, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Dog, Human
  • Immunogen

    Fusion protein corresponding to Rabbit CACNA1C aa 1507-1733 (C terminal).

  • Positive control
    • WB: Cell lysates prepared from DHPR alpha 1 transfected CHO cells. IHC-P: Human hippocampus tissue; Mouse brain tissue. ICC/IF: SK-N-BE cells. Flow Cyt: SH-SY5Y cells. IHC-Fr: Rabbit heart tissue.
  • General notes

    The clone number has been updated from S57-46 to L57/46, both clone numbers name the same antibody clone.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab84814 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
IHC-P Use a concentration of 0.1 - 10 µg/ml. Use at 0.1-1.0µg/mL (perox), 1.0-10µg/mL (IF).
IHC-Fr Use a concentration of 0.1 - 10 µg/ml. Use at 0.1-1.0µg/mL (perox), 1.0-10µg/mL (IF).
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 240 kDa. If results are poor, use lysate without boiling, heat at 37°C for 15 minutes.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Voltage-sensitive calcium channels (VSCC) mediate the entry of calcium ions into excitable cells and are also involved in a variety of calcium-dependent processes, including muscle contraction, hormone or neurotransmitter release, gene expression, cell motility, cell division and cell death. The isoform alpha-1C gives rise to L-type calcium currents. Long-lasting (L-type) calcium channels belong to the 'high-voltage activated' (HVA) group. They are blocked by dihydropyridines (DHP), phenylalkylamines, benzothiazepines, and by omega-agatoxin-IIIA (omega-Aga-IIIA). They are however insensitive to omega-conotoxin-GVIA (omega-CTx-GVIA) and omega-agatoxin-IVA (omega-Aga-IVA). Calcium channels containing the alpha-1C subunit play an important role in excitation-contraction coupling in the heart. The various isoforms display marked differences in the sensitivity to DHP compounds. Binding of calmodulin or CABP1 at the same regulatory sites results in an opposit effects on the channel function.
  • Tissue specificity
    Expressed in brain, heart, jejunum, ovary, pancreatic beta-cells and vascular smooth muscle. Overall expression is reduced in atherosclerotic vascular smooth muscle.
  • Involvement in disease
    Timothy syndrome
    Brugada syndrome 3
  • Sequence similarities
    Belongs to the calcium channel alpha-1 subunit (TC 1.A.1.11) family. CACNA1C subfamily.
  • Domain
    Each of the four internal repeats contains five hydrophobic transmembrane segments (S1, S2, S3, S5, S6) and one positively charged transmembrane segment (S4). S4 segments probably represent the voltage-sensor and are characterized by a series of positively charged amino acids at every third position.
    Binding of intracellular calcium through the EF-hand motif inhibits the opening of the channel.
  • Post-translational
    modifications
    Phosphorylation by PKA activates the channel.
  • Cellular localization
    Membrane. Cell membrane. The interaction between RRAD and CACNB2 regulates its trafficking to the cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • alpha-1 polypeptide antibody
    • cardiac muscle antibody
    • isoform 1 antibody
    • L type antibody
    • CAC1C_HUMAN antibody
    • CACH 2 antibody
    • CACH2 antibody
    • CACN 2 antibody
    • CACN2 antibody
    • CACNA1C antibody
    • CACNL1A1 antibody
    • Calcium channel antibody
    • Calcium channel cardic dihydropyridine sensitive alpha 1 subunit antibody
    • Calcium channel L type alpha 1 polypeptide isoform 1 cardiac muscle antibody
    • Calcium channel voltage dependent L type alpha 1C subunit antibody
    • CaV1.2 antibody
    • CCHL1A1 antibody
    • DHPR alpha 1 antibody
    • DHPR alpha 1 subunit antibody
    • LQT8 antibody
    • TS antibody
    • Voltage dependent L type calcium channel alpha 1C subunit antibody
    • Voltage dependent L type calcium channel subunit alpha 1C antibody
    • Voltage gated calcium channel alpha subunit Cav1.2 antibody
    • Voltage gated calcium channel subunit alpha Cav1.2 antibody
    • Voltage gated L type calcium channel Cav1.2 alpha 1 subunit, splice variant 10* antibody
    • Voltage-dependent L-type calcium channel subunit alpha-1C antibody
    • Voltage-gated calcium channel subunit alpha Cav1.2 antibody
    see all

Images

  • All lanes : Anti-CACNA1C antibody [L57/46] (ab84814) at 1 µg/ml

    Lane 1 : Molecular weight marker
    Lane 2 : Cell lysates prepared from DHPR alpha 1 transfected CHO cells
  • ab84814 staining CACNA1C in human hippocampus tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin embedded tissue sections). Tissue underwent heat mediate fixation in microwave and in citrate buffer. The primary antibody was used at 1/100 dilution. A Fluorophore conjugated goat anti-mouse was used as secondary at 1/50 dilution.

  • ab84814 staining CACNA1C in mouse brain tissue section by immunohistochemistry (Bouin's fixed paraffin embedded tissue section. Tissue underwent heat mediated fixation in microwave and in citrate buffer. The primary antibody was used at 1/100 dilution. A HRP conjugated secondary was used at 10 dilution.

  • ab84814 staining CACNA1C in rabbit heart tissue by Immunohistochemistry (Frozen sections).Tissue was fixed with paraformaldehyde, permeabilized using Triton X-100 and then incubated with ab84814 at a 1/100 dilution for 12 hours at 4°C. The secondary used was an AlexaFluor® 555 conjugated goat anti-mouse monoclonal, used at a 1/500 dilution.

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with ab84814 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab84814, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

  • Formaldehyde-fixed SK-N-BE cells stained for CACNA1C (green) using ab84814 at 1/100 dilution in ICC/IF.

    Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1/200 dilution for 60 minutes at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain (A); DAPI (B, blue) nuclear stain at 1/1000, 1/5000 for 60 minutes at room temperature, 5 minutes at room temperature.

     

References

This product has been referenced in:
  • Lai YJ  et al. Estrogen receptor a promotes Cav1.2 ubiquitination and degradation in neuronal cells and in APP/PS1 mice. Aging Cell N/A:e12961 (2019). Read more (PubMed: 31012223) »
  • Gao Y  et al. Calcium Receptor and Nitric Oxide Synthase Expression in Circular Muscle of Lower Esophagus from Patients with Achalasia. Chin Med J (Engl) 131:2882-2885 (2018). Read more (PubMed: 30511698) »
See all 12 Publications for this product

Customer reviews and Q&As

11-15 of 15 Abreviews or Q&A

Answer

Thank you for your reply.
I am sorry that the protocol tips did not prove to be successful. Under our Abpromise we do guarantee this antibody for 6 months to work in western blot on human samples. Since we have not been able to resolve this issue for you, then I would be happy to send you a replacement antibody as requested..
To do that, I need to have your original order information. I believe I have found that information, could you confirm that the product was ordered on 9th Nov 2011 by Bethann Gabris. If this information is incorrect, could you send me the correct information? Also, I can send you either ab81980 or ab81095, just let me know which one you would like to receive.
I look forward to your reply and resolving this issue for you.

Read More

Answer

Thank you for contacting Abcam. I am sorry that you are having problems with ab84814 in western blot. I have looked over the images and protocol information that you sent and there are a few protocol changes that I would suggest that may help to reduce the background that you are seeing: 1 - Switch to a 5% BSA block rather than 5% Milk, as BSA is a little better a blocking non-specific binding. 2 - Increase the number of wash steps you are doing (up 5 washes, 5 minutes each) and add 0.05% Tween 20 to the wash buffer, if you have not done so already. 3 - After the secondary incubation, wash the membrane for 30 minutes in a 0.5M NaCl. Then rinse the membrane thoroughly in dH20 and develop as normal. If these protocol tips do not work then I would be happy to send you a replacement antibody, as under our Abpromise we do guarantee this antibody to work in western blot on human samples. I look forward to your reply and helping you resolve these issues.

Read More

Answer

Thank you for contacting Abcam. Under our Abpromise we guarantee all of our antibodies to work as stated on the datasheet, if they do work as stated and we cannot offer any assistance to resolve the matter, then we would either replace the antibody or give a refund. Both ab81980 and ab84814 are guaranteed to work in western blot on human samples. I would recommended ab84814, even though the western blot image may not be the cleanest. If there is anything else I can help you with, please let me know.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Rabbit Tissue sections (rabbit heart)
Specification
rabbit heart
Fixative
Paraformaldehyde
Permeabilization
Yes - triton-X 100

Abcam user community

Verified customer

Submitted Sep 29 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hESC-derived cardiomyocyte)
Specification
hESC-derived cardiomyocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 03 2010

11-15 of 15 Abreviews or Q&A

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