Overview

  • Product name

    Anti-Calcineurin A antibody
    See all Calcineurin A primary antibodies
  • Description

    Rabbit polyclonal to Calcineurin A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Chicken, Cow, Dog, Turkey, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
  • Immunogen

    A synthetic peptide corresponding to a region between residue 470 and the C-terminus (residue 521) of human Calcineurin A (NP_000935.1).

  • Positive control

    • Whole cell lysate from hela cells, NIH3T3 cells and 293T cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab71149 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
WB 1/2000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 59 kDa).
IP Use at 2-5 µg/mg of lysate.
IHC-P 1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Calcium-dependent, calmodulin-stimulated protein phosphatase. This subunit may have a role in the calmodulin activation of calcineurin. Dephosphorylates DNM1L, HSPB1 and SSH1.
  • Sequence similarities

    Belongs to the PPP phosphatase family. PP-2B subfamily.
  • Cellular localization

    Nucleus. Colocalizes with ACTN1 and MYOZ2 at the Z line in heart and skeletal muscle.
  • Information by UniProt
  • Database links

  • Alternative names

    • Alpha isoform formerly PPP2B antibody
    • Calcineurin A alpha antibody
    • Calcineurin A1 antibody
    • CalcineurinA antibody
    • Calmodulin dependent calcineurin A subunit alpha isoform antibody
    • Calmodulin-dependent calcineurin A subunit alpha isoform antibody
    • CALN antibody
    • CALNA 1 antibody
    • CALNA antibody
    • CALNA1 antibody
    • CAM PRP catalytic subunit antibody
    • CAM-PRP catalytic subunit antibody
    • CCN 1 antibody
    • CCN1 antibody
    • CNA 1 antibody
    • CNA alpha antibody
    • CNA antibody
    • CNA1 antibody
    • PP2BA_HUMAN antibody
    • PPP2B antibody
    • Ppp3ca antibody
    • Protein phosphatase 2B catalytic subunit antibody
    • Protein phosphatase 3 (formerly 2B) catalytic subunit alpha isoform antibody
    • Protein phosphatase 3 catalytic subunit alpha isoform PPP3CA antibody
    • Protein phosphatase 3 catalytic subunit alpha isozyme antibody
    • Serine/threonine protein phosphatase 2B catalytic subunit alpha isoform antibody
    • Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform antibody
    see all

Images

  • All lanes : Anti-Calcineurin A antibody (ab71149) at 0.1 µg/ml

    Lane 1 : Whole cell lysate from Hela cells at 50 µg
    Lane 2 : Whole cell lysate from Hela cells at 15 µg
    Lane 3 : Whole cell lysate from Hela cells at 5 µg
    Lane 4 : Whole cell lysate from 293T cells at 50 µg
    Lane 5 : Whole cell lysate from NIH 3T3 cells at 50 µg

    Predicted band size: 59 kDa
    Observed band size: 59 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling Calcineurin A with ab71149 at 1/1000 (1µg/ml). Detection: DAB.
  • Immunoprecipitation/ Western Blot of Calcineurin A
    Lane 1: ab71149 at 3µg/mg whole cell lysate.
    Lane 2: Control IgG.
    Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
    Subsequent WB detection was performed with 1 µg/ml ab71149.
    Chemiluminescence with an exposure time of 30 seconds.

References

This product has been referenced in:

  • Haryuna TS  et al. Curcumin Reduces the Noise-Exposed Cochlear Fibroblasts Apoptosis. Int Arch Otorhinolaryngol 20:370-376 (2016). IHC ; Rat . Read more (PubMed: 27746842) »
  • Thoms KM  et al. Cyclosporin A, but not everolimus, inhibits DNA repair mediated by calcineurin: implications for tumorigenesis under immunosuppression. Exp Dermatol 20:232-6 (2011). WB ; Human . Read more (PubMed: 21323745) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your reply. This antibody is specific for the calcineurin A alpha isoform: http://www.uniprot.org/uniprot/Q08209 I hope this information helps. Please contact us with any other questions.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (GM00637 fibroblast cell line)
Loading amount
50 µg
Specification
GM00637 fibroblast cell line
Gel Running Conditions
Reduced Denaturing (10 %)
Blocking step
(agent) for 30 minute(s) · Concentration: 5µg/mL · Temperature: 20°C

Mrs. Annika Schäfer

Verified customer

Submitted Apr 09 2009

Answer

Thank you for your inquiry. We have not seen a banding pattern similar to what you're describing with in-house testing. We have predominantly seen a single band at 60 kDa. However, we have heard from a customer who saw something similar to what you're describing using fibroblast cell lysate. https://www.abcam.com/index.html?pageconfig=reviews&intAbID=71149&strFilterApplication=Western blot Important to note is that this protein does have two isoforms, at 57 and 59 kDa, respectively. There are a few things that may be going on here, including degradation of 1 isoform, resolution of the isoforms in an unexpected way due to experimental variances in gel percentage, running buffers, etc. Therefore, I would be inclined to think that both bands are specific to calcineurin A. You may want to run a positive control with HeLa, 293T, or NIH 3T3 cells. Also, take a look at this reference which used this antibody in WB to make sure your protocol is similar: http://www.ncbi.nlm.nih.gov/pubmed/21323745?dopt=Abstract Below is the protocol that was tested in-house: Reagents Needed: 20X Running Buffer Tricine (free base) 71.7 g Tris (free base) 72.6 g SDS 10.0 g Sodium Bisulfite 2.5 g Adjust to 500 ml with ultra pure water. Store at 4°C. For 1X Running Buffer, add 10 ml of 20X Running Buffer to 190 ml of distilled water. 10X Transfer Buffer Tris (free base) 15.2 g Glycine 72.1 g SDS 5.0 g Ultra pure water to 500 ml 10X Transfer Buffer Store at 4°C. 1X Transfer Buffer 10X Transfer Buffer 50 ml Methanol 100 ml Distilled water 350 ml Make fresh for each use. 5% non-fat dry milk in TBST Carnation non-fat dry milk 50 g TBST 1 liter TBST (Tris Buffered Saline with Tween 20, pH8.0) Tris 6.1 g NaC l 8.68 g Tween-20 500 mcl Adjust the volume to 1 liter with distilled water. Adjust pH to 8.0 with HCL. Store at 4-25°C. Electrophoresis in SDS-Polyacrylamide Gel and Transfer to Nitrocellulose: Polyacrylamide gels (4-12% Tricine), running buffer and transfer buffer SeeBlue2 and HiMark molecular weight markers Nitrocellulose membranes Cut open the package that contains the gel cassette and drain away the buffer. Rinse the wells with distilled water. Rinse the wells with fresh 1x running buffer. Place the gels on the buffer core so that the shorter plates face inward. If only using one gel, use a buffer dam to seal the other side. Fill the inner core with fresh 1X running buffer. If there are no leaks, fill the outer core with running buffer. Load samples. Run the gels at 150V until the dye front reaches the bottom of the gel (approximately 60 minutes). Soak nitrocellulose membrane and blotting paper in 1X transfer buffer for at least 5 minutes prior to opening gel cassette. Open gel cassettes and place the gel on the nitrocellulose membrane sandwiched between two pieces of blotting paper. Place in transfer apparatus and fill with fresh 1X transfer buffer. Run transfer apparatus for 60-75 minutes on 35V. Western Blotting: Remove the membrane from the transfer apparatus and place in 20 ml of 5% non-fat dry milk in TBST for one hour, with gentle shaking. Dilute the primary antibody in 15 ml of 5% non-fat dry milk in TBST. For best results, the optimal dilution of antibody should be empirically defined. Incubate the membrane in diluted primary antibody for two hours to overnight with gentle shaking at room temperature. Wash the membrane three times, 10 minutes each time in TBST. Dilute the secondary HRP conjugated antibody in 15 ml of reconstituted 5% non-fat dry milk in TBST. For best results, the optimal concentration of the secondary HRP conjugated antibody should be empirically defined. Incubate the membrane in diluted anti-Rabbit IgG-HRP Conjugate for 60 minutes. Wash as directed in step 4. Develop blots with substrate solution and place in plastic membrane protector. Expose membrane to film or CCD camera. I hope this information helps. If you're still having difficulty, please let me know and I will be happy to help you further.

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