Caldesmon Recombinant Rabbit Monoclonal Antibody [SU03-19]
cat.: ET1608-16
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SU03-19
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 93 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Caldesmon aa 744-793 / 793.
Positive control: NIH/3T3 cell lysates, NIH/3T3, C6, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location: Cytoplasm, Cytoskeleton.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q05682 Human | Q62736 Rat
Unigene: 308134 Mouse
Alternative names: CAD CALD 1 CALD1 CALD1_HUMAN Caldesmon 1 Caldesmon 1 Isoform 1 Caldesmon 1 Isoform 2 Caldesmon 1 Isoform 3 Caldesmon 1 Isoform 4 Caldesmon 1 Isoform 5 Caldesmon Caldesmon1 CDM H CAD HCAD L CAD LCAD MGC21352 NAG22
Images
ET1608-16_1.jpg Fig1: Western blot analysis of Caldesmon on NIH/3T3 cell lysates with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 93 kDa
Observed band size: 93 kDa

Exposure time: 2 minutes;

8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-16) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1608-16_2.jpg Fig2: Immunocytochemistry analysis of NIH/3T3 cells labeling Caldesmon with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1608-16_3.jpg Fig3: Immunocytochemistry analysis of C6 cells labeling Caldesmon with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1608-16_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-16) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-16_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-16) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-16_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Caldesmon antibody (ET1608-16) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-16) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-16_7.jpg Fig7: Flow cytometric analysis of Caldesmon was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-16, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
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