Overview

  • Product name
  • Description
    Rabbit polyclonal to FKBP12
  • Specificity
    Detects FK506 binding protein 12 kDa (FKBP12).
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IP, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Pig
    Predicted to work with: Rabbit, Cow, Xenopus laevis
  • Immunogen

    Synthetic peptide corresponding to Human FKBP12 aa 1-13 (N terminal).
    Sequence:

    GVQVETISPGDGR


    (Peptide available as ab4935)

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    Immunophilins are a family of soluble receptors capable of binding to one of two major immunosuppressant agents; cyclosporin A (CsA) or FK506. Proteins that bind FK506 are termed FK506 Binding Proteins (FKBPs) and those that bind cyclosporin A are called cyclophilins (CyP). Immunophilins function as cis-trans peptidyl-prolyl isomerases (PPIase) whose activity is inhibited by their respective immunosuppressant compounds. Thus, immunophilins accelerate folding of some proteins both in vivo and in vitro by catalyzing slow steps in the initial folding and rearrangement of proline-containing proteins. FKBP12:FK506 complexes inhibit calcineurin, a calcium/calmodulin-dependent serine/threonine phosphatase which blocks T-cell activation by preventing lymphokine gene transcription. FKBP12 also plays a role in intracellular calcium regulation by associating with three types of calcium release channel complexes, cardiac and skeletal ryanodine receptors and the inositol 1,4,5-trisphosphate receptor. In interactions with members of the TGF beta family type I receptors, FKBP12 has been shown to exert an inhibitory effect on the signaling pathway.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2918 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
IHC-P 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Can be blocked with FKBP12 peptide (ab4935).
IHC-Fr 1/700. PubMed: 16171515

Target

  • Function
    May play a role in modulation of ryanodine receptor isoform-1 (RYR-1), a component of the calcium release channel of skeletal muscle sarcoplasmic reticulum. There are four molecules of FKBP12 per skeletal muscle RYR. PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
  • Sequence similarities
    Belongs to the FKBP-type PPIase family. FKBP1 subfamily.
    Contains 1 PPIase FKBP-type domain.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • 12 kDa FK506-binding protein antibody
    • 12 kDa FKBP antibody
    • Calstabin 1 antibody
    • FK506 binding protein 1 antibody
    • FK506 binding protein 12 antibody
    • FK506 binding protein 1A (12kD) antibody
    • FK506 binding protein 1A 12kDa antibody
    • FK506 binding protein 1A antibody
    • FK506 binding protein T cell 12 kD antibody
    • FK506 binding protein, T cell, 12 kD antibody
    • FK506 binding protein12 antibody
    • FK506-binding protein 1A antibody
    • FKB1A_HUMAN antibody
    • FKBP 12 antibody
    • FKBP 1A antibody
    • FKBP-12 antibody
    • FKBP-1A antibody
    • FKBP1 antibody
    • FKBP12 antibody
    • FKBP12 Exip3 antibody
    • FKBP12C antibody
    • fkbp1a antibody
    • Immunophilin FKBP12 antibody
    • Peptidyl prolyl cis trans isomerase antibody
    • Peptidyl-prolyl cis-trans isomerase FKBP1A antibody
    • PKC12 antibody
    • PKCI2 antibody
    • PPIase antibody
    • PPIase FKBP1A antibody
    • Protein kinase C inhibitor 2 antibody
    • Rotamase antibody
    see all

Images

  • All lanes : Anti-FKBP12 antibody (ab2918) at 1/1000 dilution

    Lane 1 : Whole tissue lysate, dog left ventricle (heart failure)
    Lane 2 : Whole tissue lysate, dog left ventricle (heart failure)
    Lane 3 : Whole tissue lysate, dog left ventricle (heart failure)
    Lane 4 : Whole tissue lysate, dog left ventricle (heart failure)
    Lane 5 : Whole tissue lysate, dog left ventricle (heart failure)
    Lane 6 : Whole tissue lysate, dog left ventricle (normal)
    Lane 7 : Whole tissue lysate, dog left ventricle (normal)
    Lane 8 : Whole tissue lysate, dog left ventricle (normal)
    Lane 9 : Whole tissue lysate, dog left ventricle (normal)

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP conjugated donkey polyclonal antibody
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 12 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an Abreview submitted by Dr sudhish mishra

    See Abreview

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/50 with a rabbit polyclonal antibody recognizing FKBP12 (ab2918) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • ICC/IF image of ab2918 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2918, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human colon carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing FKBP12 (ab2918) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/20 with a rabbit polyclonal antibody recognizing FKBP12 (ab2918) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • ab2918 at 1/1000 staining FKBP12 from human penis tissue by IHC-P. The tissue was paraformaldehyde fixed and the slides were incubated in citrate buffer for antigen retrieval and heated for 20 minutes. The tissue was incubated with ab2918 for 24 hours. The secondary antibody used was part of the DAKO ENVISON System. The picture depicts staining of nerve fibers within corporal tissue of the human penis.

    See Abreview

  • ab2918 at 1/1000 staining mouse heart tissue sections by IHC-P. The tissue was paraformaldehyde fixed, blocked and then a citrate buffer / heat mediated antigen retireival step was performed. The tissue was incubated with the antibody for 24 hours. An HRP conjugated rabbit polyclonal antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Schreiber KH  et al. Rapamycin-mediated mTORC2 inhibition is determined by the relative expression of FK506-binding proteins. Aging Cell 14:265-73 (2015). WB . Read more (PubMed: 25652038) »
  • Ballister ER  et al. Recruitment of Mad1 to metaphase kinetochores is sufficient to reactivate the mitotic checkpoint. J Cell Biol 204:901-8 (2014). Read more (PubMed: 24637323) »

See all 8 Publications for this product

Customer reviews and Q&As

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Cultured mESC cells)
Total protein in input
2000 µg
Immuno-precipitation step
Protein A
Specification
Cultured mESC cells
Username

Abcam user community

Verified customer

Submitted Jun 05 2017

Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MOPS buffer))
Loading amount
75000 cells
Specification
Chinese hamster ovary (CHO) cells
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 17 2017

Application
Western blot
Sample
Rabbit Cell lysate - whole cell (Rabbit Reticulocyte)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MES buffer))
Loading amount
0.5 µg
Specification
Rabbit Reticulocyte
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 05 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Total protein in input
1e+006 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Canine Kidney Epithelial MDCK cells
Username

Abcam user community

Verified customer

Submitted Nov 04 2016

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Mouse embryonic fibroblasts (MEFs)
Username

Abcam user community

Verified customer

Submitted Nov 04 2016

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
A549 (human lung epithelium cells)
Username

Abcam user community

Verified customer

Submitted Nov 01 2016

Application
Western blot
Sample
Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
500000 cells
Specification
Canine Kidney Epithelial MDCK cells
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 27 2016

Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (VERO E6)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
60000 cells
Specification
VERO E6
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 11 2016

Application
Western blot
Sample
Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
50000 cells
Specification
BHK-21 (Kidney Fibroblasts)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 10 2016

1-10 of 23 Abreviews or Q&A

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