Application
Immunostaining of GAPDH in A-10 cell line (rat aortic smooth muscle cells). Cells were fixed by formalin and GAPDH was stained by: (green colour) F-actin microfilaments-bindingdye (red) DNA-binding dye (dark blue)Application
Immunoprecipitation of GAPDH from rat heart extract using anti-GAPDH MAb 6C5 (A). Mixture of protein A-Sepharose with anti-GAPDH MAbs and tissue extract was incubated for 30 min at room temperature and precipitated by centrifugation. Pellet was washed with PBS, suspended in reducing electrophoresis sample buffer and heated for 5 minutes at 100 ºC. After centrifugation supernatant was loaded on gel and proteins were separated by SDS electrophoresis. Lane 1: Human GAPDH (1 µg) Lane 2: GAPDH immunoprecipitated from rat heart tissue extract Lane 3: Only preincubated with Protein A Sepharose Lane 4: Only Protein A SepharoseApplication
Heart extracts were prepared from different animal species. About 0.5 mg of homogenized wet cardiac tissue per track was loaded. Lane 1: isolated human GAPDH, 0.5 µg Lane 2: human heart tissue extract Lane 3: pig heart tissue extract Lane 4: goat heart tissue extract Lane 5: bovine heart tissue extract Lane 6: dog heart tissue extract Lane 7: mouse heart tissue extract Lane 8: rat heart tissue extract Lane 9: rabbit heart tissue extract| Product Name | GAPDH antibody [6C5] |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal [6C5] to GAPDHGlyceraldehyde-3-phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. Besides its functioning as a glycolytic enzyme in cytoplasm, recent evidence suggest that mammalian GAPDH is also involved in a great number of intracellular processes such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. During the last decade a lot of findings appeared concerning the role of GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age-related neuronal diseases, such as Alzheimer's and Huntington's disease. GAPDH is constitutively expressed in almost all tissues at high levels, therefore becoming the marker of choice when a loading control in Western blotting is required. Some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types. |
| Immunogen | Rabbit muscle GAPDH. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Clone Number | 6C5 |
| Host Species | Mouse |
| Tested Applications | PurifyCell CultureControlELISAICC/IFIPWB |
| AP: Use at an assay dependent dilution. ELISA: Use at an assay dependent dilution. ICC: Use at an assay dependent dilution. WB: Use at a concentration of 1 - 10 μg/ml. Gives a very strong band in Western Blotting in concentration 10 μg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). Optimal dilutions/concentrations should be determined by the end user. | |
| Species Reactivity | HumanMouseDogPigRatCatFishRabbit |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Synonyms | G3PD G3PDH GAPD Glyceraldehyde 3 phosphate dehydrogenase Glyceraldehyde 3 phosphate dehydrogenase liver Glyceraldehyde 3 phosphate dehydrogenase muscle |
|---|---|
| Target | GAPDH |
| Tissue Specificity | This antibody can be used as a loading control antibody. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. There is no cross-reaction with GAPDH from yeast. |
| Cellular Localization | Cytoplasmic |
Application
Immunostaining of GAPDH in A-10 cell line (rat aortic smooth muscle cells). Cells were fixed by formalin and GAPDH was stained by: (green colour) F-actin microfilaments-bindingdye (red) DNA-binding dye (dark blue)
Application
Immunoprecipitation of GAPDH from rat heart extract using anti-GAPDH MAb 6C5 (A). Mixture of protein A-Sepharose with anti-GAPDH MAbs and tissue extract was incubated for 30 min at room temperature and precipitated by centrifugation. Pellet was washed with PBS, suspended in reducing electrophoresis sample buffer and heated for 5 minutes at 100 ºC. After centrifugation supernatant was loaded on gel and proteins were separated by SDS electrophoresis. Lane 1: Human GAPDH (1 µg) Lane 2: GAPDH immunoprecipitated from rat heart tissue extract Lane 3: Only preincubated with Protein A Sepharose Lane 4: Only Protein A Sepharose
Application
Heart extracts were prepared from different animal species. About 0.5 mg of homogenized wet cardiac tissue per track was loaded. Lane 1: isolated human GAPDH, 0.5 µg Lane 2: human heart tissue extract Lane 3: pig heart tissue extract Lane 4: goat heart tissue extract Lane 5: bovine heart tissue extract Lane 6: dog heart tissue extract Lane 7: mouse heart tissue extract Lane 8: rat heart tissue extract Lane 9: rabbit heart tissue extract| Application Notes | AP: Use at an assay dependent dilution. ELISA: Use at an assay dependent dilution. ICC: Use at an assay dependent dilution. WB: Use at a concentration of 1 - 10 μg/ml. Gives a very strong band in Western Blotting in concentration 10 μg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). Optimal dilutions/concentrations should be determined by the end user. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution, aliquot and store at 4°C. Do not freeze. We recommend to parafilm vials during storage to prevent evaporation of buffer. |
| Storage Buffer | Phosphate-buffered saline, pH 7.4, containing 0.1% sodium azide |
