Anti-PCK2 antibody
| Name | Anti-PCK2 antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab70359 |
| Prices | $385.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | ICC/IF ICC/IF IHC-P WB IP |
| Species Reactivities | Rat, Human, Mouse, Pig |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human PCK2 |
| Description | Rabbit Polyclonal |
| Gene | PCK2 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Anti-PCK2 antibody (ab70359) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µgSecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![PCK2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to PCK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab70359.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 71kDa: PCK2.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/8390_PCK2-Primary-antibodies-ab70359-7.jpg)
PCK2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to PCK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab70359.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 71kDa: PCK2.
ICC/IF image of ab70359 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab70359, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and Hek293 cells at 5µg/ml.
IHC image of PCK2 staining in normal human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab70659, 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.
Product References
Mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M) is a pro-survival, - Mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M) is a pro-survival,
Mendez-Lucas A, Hyrossova P, Novellasdemunt L, Vinals F, Perales JC. J Biol Chem. 2014 Aug 8;289(32):22090-102.
PEPCK-M expression in mouse liver potentiates, not replaces, PEPCK-C mediated - PEPCK-M expression in mouse liver potentiates, not replaces, PEPCK-C mediated
Mendez-Lucas A, Duarte JA, Sunny NE, Satapati S, He T, Fu X, Bermudez J, Burgess SC, Perales JC. J Hepatol. 2013 Jul;59(1):105-13.