Overview

  • Product name
    Anti-GM130 antibody [EP892Y] - BSA and Azide free
    See all GM130 primary antibodies
  • Description
    Rabbit monoclonal [EP892Y] to GM130 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IP, ICC/IF, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Cow, Dog, Human, Monkey, African green monkey
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human GM130 aa 1-100.

  • Positive control
    • WB: HeLa, MCF7, MDCK(NBL-2), MDBK(BL-1) and COS-1 cell lysates. IHC-P: Human cervix carcinoma and liver tissues. ICC/IF: HeLa and MCF7 cells.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab215966 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
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

PFA fixation should be most suitable.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Overnight incubation is recommended.

WB Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 112 kDa).
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Golgi auto-antigen; probably involved in maintaining cis-Golgi structure.
  • Sequence similarities
    Belongs to the GOLGA2 family.
  • Domain
    Extended rod-like protein with coiled-coil domains.
  • Cellular localization
    Golgi apparatus > Golgi stack membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 130 kDa cis Golgi matrix protein antibody
    • 130 kDa cis-Golgi matrix protein antibody
    • Cis golgi matrix protein GM130 antibody
    • GM130 antibody
    • Gm130 autoantigen antibody
    • GOGA2_HUMAN antibody
    • GOLGA 2 antibody
    • Golga2 antibody
    • Golgi autoantigen antibody
    • Golgi autoantigen golgin subfamily a 2 antibody
    • Golgi matrix protein GM130 antibody
    • Golgin 95 antibody
    • golgin A2 antibody
    • Golgin subfamily a 2 antibody
    • Golgin subfamily A member 2 antibody
    • Golgin-95 antibody
    • MGC20672 antibody
    • SY11 protein antibody
    see all

Images

  • Effect of Irgm1 palmitoylation mutation on Golgi association

    Irgm1 KO MEF were transfected with plasmids expressing wild-type or mutant Irgm1 proteins, as indicated. The cells were exposed to 100 U/ml IFN-γ for 24 h, stained with anti-Irgm1 and anti-GM130 antibodies, and used for immunofluorescence analysis. The experiment was performed 3 times, with at least 20 cells analyzed per group in each experiment. (A) Shown are images from representative cells. The scale bar represents 20 µm. 

    Cells are 4% paraformaldehyde fixed, 0.2% saponin-permeabilized.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling GM130 (red) with ab52649 at a 1/20 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling GM130 with purified ab52649 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling GM130 with purified ab52649 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/50) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • ab52649 (purified) at 1/20 immunoprecipitating GM130 in HeLa whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate (10µg)

    Lane 2 (+): ab52649 + HeLa whole cell lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52649 in HeLa whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Unpurified ab52649 staining GM130 (magenta) in monkey kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 3% BSA + 0.5% Triton X-100 for 45 minutes at 25°C. Samples were incubated with primary antibody (1/1500 in 3% BSA + 0.5% Triton X-100) for 45 minutes at 25°C. An Alexa Fluor® 647-conjugated donkey anti-rabbit IgG polyclonal (2 µg/ml) was used as the secondary antibody. Nuclei stained with Picogreen.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling GM130 with unpurified ab52649 at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Unpurified ab52649 staining GM130 in human ARPE-19 cells by ICC/IF (immunocytochemistry/immunofluorescence). Cells were formaldehyde fixed, permeabilized by 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. The sample was incubated with the primary antibody (1/500 in 1% goat serum, 0.1%TX100, 1 x PBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit polyclonal (1/500) was used as the secondary.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • Unpurified ab52649 staining GM130 in Bovine brain microvascular endothelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% saponin and blocked with 5% BSA for 90 minutes at 37°C. Samples were incubated with primary antibody (1/100 in 0.1% saponin + 1% BSA ) for 18 hours at 4°C. An undiluted Alexa Fluor® 568-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

  • ICC/IF image of unpurified ab52946 stained HeLa cells. The cells were 4% PFA 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 (unpurified ab52946, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52649).

References

This product has been referenced in:
  • Raza S  et al. A bovine herpesvirus 1 pUL51 deletion mutant shows impaired viral growth in vitro and reduced virulence in rabbits. Oncotarget N/A:N/A (2016). Read more (PubMed: 26934330) »
  • Mazzulli JR  et al. a-Synuclein-induced lysosomal dysfunction occurs through disruptions in protein trafficking in human midbrain synucleinopathy models. Proc Natl Acad Sci U S A 113:1931-6 (2016). Read more (PubMed: 26839413) »
See all 26 Publications for this product

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