Overview

  • Product name

  • Description

    Rabbit polyclonal to PGAM5
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human, Zebrafish
    Predicted to work with: Mouse, Rat, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human PGAM5.

    Read Abcam's proprietary immunogen policy

  • Positive control

    • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
    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...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • Function

    Displays 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 similarities

    Belongs to the phosphoglycerate mutase family. BPG-dependent PGAM subfamily.
  • Domain

    According 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 localization

    Mitochondrion. 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
  • Database links

  • Alternative names

    • Bcl-XL-binding protein v68 antibody
    • BXLBv68 antibody
    • MGC5352 antibody
    • mitochondrial antibody
    • PGAM5 antibody
    • PGAM5_HUMAN antibody
    • Phosphoglycerate mutase family member 5 antibody
    • Serine/threonine protein phosphatase PGAM5 mitochondrial antibody
    • Serine/threonine-protein phosphatase PGAM5 antibody
    see all

Images

  • 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.

    Secondary
    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.

References

ab131552 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Tissue lysate - whole (HELA cells)
Gel Running Conditions
Non-reduced Denaturing (10% Bis-Tris MOPS gel)
Loading amount
20 µg
Specification
HELA cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Ventana Ultra CC1
Permeabilization
No
Specification
Tonsil
Blocking step
Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 01 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Zebrafish Tissue lysate - whole (total larvae, 4 days post fertilization)
Specification
total larvae, 4 days post fertilization
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Dr. Evgenia Salta

Verified customer

Submitted Oct 27 2014

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