WB result of HDAC6 Rabbit mAb
Primary antibody: HDAC6 Rabbit mAb at 1/1000 dilution
Lane 1: A549 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 126 kDa
Observed MW: 160 kDa
(This blot was developed with high sensitivity substrate)
S-RMab®HDAC6 Recombinant Rabbit mAb (S-R338)
S-RMab®HDAC6 Recombinant Rabbit mAb (S-R338)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | HDAC6 |
| Synonyms | Histone deacetylase 6, HD6, Histone deacetylase mHDA2, Tubulin-lysine deacetylase HDAC6 |
| Location | Cytoplasm, Nucleus |
| Accession | Q9Z2V5 |
| Clone Number | S-R338 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IHC-P | 1:500 | |
| ICC | 1:100 | |
| ICFCM | 1:50 |
Background
Histone deacetylase 6 is an enzyme that in humans is encoded by the HDAC6 gene. HDAC6 belongs to class II of the histone deacetylase/acuc/apha family. It contains an internal duplication of two catalytic domains that appear to function independently of each other. This protein possesses histone deacetylase activity and represses transcription. Since HDAC6 is dysregulated and/or implicated in several cancers and neurodegenerative disorders, pharmacological inhibition of this specific enzyme holds great therapeutic potential and could also limit side effects associated with pan-inhibitors of multiple HDAC enzymes.
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized A549 (Human lung carcinoma epithelial cell) labelling HDAC6 antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti-HDAC6 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-HDAC6 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung adenocarcinoma. Anti-HDAC6 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows clustered cytoplasm staining in A549 cells, treated with MG132 (5 μM, 24 hr). Anti-HDAC6 antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
Negative control: ICC shows homogeneous cytoplasm staining in A549 cells, untreated with MG132 (5 μM, 24 hr). Anti-HDAC6 antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
