Phospho-Vimentin (S39) Recombinant Rabbit Monoclonal Antibody [JE43-26]
cat.: HA721442
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | JE43-26 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. 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: 54 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues surrounding Ser39 of human vimentin protein. |
| Positive control: | HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate, HeLa treated with 100nM Calyculin A for 30 minutes, NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, C6 treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Cytoplasm, cytoskeleton, Nucleus matrix, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell |
1:1,000 1:100 |
| Uniprot #: | SwissProt: P08670 Human | P20152 Mouse | P31000 Rat |
| Alternative names: | CTRCT30 Epididymis luminal protein 113 FLJ36605 HEL113 VIM VIME_HUMAN Vimentin |
Images
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Fig1:
Western blot analysis of Phospho-Vimentin (S39) on different lysates with Rabbit anti-Phospho-Vimentin (S39) antibody (HA721442) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time: 10 seconds; 4-20% 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 (HA721442) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Western blot analysis of Phospho-Vimentin (S39) on different lysates with Rabbit anti-Phospho-Vimentin (S39) antibody (HA721442) at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: C6 cell lysate Lane 4: C6 treated with 100nM Calyculin A for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time: 30 seconds; 4-20% 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 (HA721442) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Immunocytochemistry analysis of HeLa cells treated with or without 100nM Calyculin A for 30 minutes, then treated with λpp for 1 hour labeling Phospho-Vimentin (S39) with Rabbit anti-Phospho-Vimentin (S39) antibody (HA721442) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-Vimentin (S39) antibody (HA721442) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, 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. |
|
Fig4:
Immunocytochemistry analysis of NIH/3T3 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-Vimentin (S39) with Rabbit anti-Phospho-Vimentin (S39) antibody (HA721442) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Phospho-Vimentin (S39) antibody (HA721442) 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 (M1305-2, 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.