Product nameAnti-PGAM5 antibody
See all PGAM5 primary antibodies
DescriptionRabbit polyclonal to PGAM5
Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
Species reactivityReacts with: Human, Zebrafish
Predicted to work with: Mouse, Rat, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster
Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human PGAM5.
- This antibody gave a positive signal in HepG2 whole cell lysate as well as the following Human tissue lysates: Brain; Liver; Tonsil. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal pancreas. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab131552 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 32, 35 kDa (predicted molecular weight: 32 kDa). Abcam recommends using milk as the blocking agent - 3%.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionDisplays phosphatase activity for serine/threonine residues, and, dephosphorylates and activates MAP3K5 kinase. Has apparently no phosphoglycerate mutase activity. May be regulator of mitochondrial dynamics. Substrate for a KEAP1-dependent ubiquitin ligase complex. Contributes to the repression of NFE2L2-dependent gene expression.
Sequence similaritiesBelongs to the phosphoglycerate mutase family. BPG-dependent PGAM subfamily.
DomainAccording to PubMed:18387606, may contain a non-cleaved N-terminal mitochondrial targeting sequence that targets PGAM5 to the cytosolic side of the outer mitochondrial membrane, instead of a N-terminal transmembrane.
Cellular localizationMitochondrion. Mitochondrion outer membrane. Membrane. Isoform 2 overexpression results in the formation of disconnected punctuate mitochondria distributed throughout the cytoplasm. Isoform 1 overexpression results in the clustering of mitochondria around the nucleus.
- Information by UniProt
- Bcl-XL-binding protein v68 antibody
- BXLBv68 antibody
- MGC5352 antibody
All lanes : Anti-PGAM5 antibody (ab131552) at 1 µg/ml (Milk block - 3%)
Lane 1 : Human liver tissue lysate - total protein (ab29889)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Tonsil (Human) Tissue Lysate
Lane 4 : Human brain tissue lysate - total protein (ab29466)
Lysates/proteins at 25 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDa
Additional bands at: 35 kDa, 73 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
Abcam recommends using milk as the blocking agent - 3%. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab131552 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
ICC/IF image of ab131552 stained HeLa 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 ab131552 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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.
IHC image of PGAM5 staining in Human normal pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab131552, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab131552 has not yet been referenced specifically in any publications.