Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (ab52649)

Overview

  • Product nameAnti-GM130 antibody [EP892Y] - cis-Golgi Marker
    See all GM130 primary antibodies
  • Description
    Rabbit monoclonal [EP892Y] to GM130 - cis-Golgi Marker
  • SpecificityMouse and rat cell lines pc12, 3t3, raw 264.7 were tested positive in WB. However, brain, kidney, spleen and heart were negative from the two species.
  • Tested applicationsSuitable for: ICC/IF, IHC-P, IHC-Fr, WB, IPmore 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

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-GM130 antibody (Alexa Fluor® 647) - cis-Golgi Marker [EP892Y] (ab195303)

    Anti-GM130 antibody (Alexa Fluor® 568) - cis-Golgi Marker [EP892Y]  (ab202534)

    Anti-GM130 antibody (Alexa Fluor® 594) - cis-Golgi Marker [EP892Y] (ab203433)

    Anti-GM130 antibody (Alexa Fluor® 555) - cis-Golgi Marker [EP892Y] (ab203436)        

    Anti-GM130 antibody (Alexa Fluor® 488) - cis-Golgi Marker  [EP892Y]  (ab195302)

Properties

Applications

Our Abpromise guarantee covers the use of ab52649 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 1/50 - 1/250.

PFA fixation should be most suitable.

IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Overnight incubation is recommended.

IHC-Fr Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 112 kDa).
IP 1/20 - 1/50.

Target

  • FunctionGolgi auto-antigen; probably involved in maintaining cis-Golgi structure.
  • Sequence similaritiesBelongs to the GOLGA2 family.
  • DomainExtended rod-like protein with coiled-coil domains.
  • Cellular localizationGolgi 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

Anti-GM130 antibody [EP892Y] - cis-Golgi Marker images

  • All lanes : Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (ab52649) at 1/5000 dilution (purified)

    Lane 1 : MDCK(NBL-2) cell lysate
    Lane 2 : MDCK(BL-1) cell lysate
    Lane 3 : COS-1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 112 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (ab52649) at 1/1000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 112 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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).

  • 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.

  • Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (ab52649) at 1/200000 dilution (unpurified) + HeLa cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 112 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling GM130 with unpurified ab52649 at a dilution of 1/500.

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

  • ICC/IF image of unpurified ab52649 stained MCF7 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 ab52649, 1µg/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.

References for Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (ab52649)

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

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HT-29)
Permeabilization Yes - PBS + 0.05% saponin
Specification HT-29
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 23 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Hippocampal neuron, 18DIV)
Gel Running Conditions Reduced Denaturing (9% SDS-PAGE)
Loading amount 10 µg
Specification Hippocampal neuron, 18DIV
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Dr. Junmo Hwang

Verified customer

Submitted Jul 14 2016

Application Immunocytochemistry
Sample Rat Cultured Cells (primary neuronal culture_Hippocampal neuron, DIV16)
Permeabilization Yes - 0.2% Triton X-100 in PBS
Specification primary neuronal culture_Hippocampal neuron, DIV16
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Fixative Paraformaldehyde
Username

Dr. Junmo Hwang

Verified customer

Submitted Apr 27 2016

Application Western blot
Sample Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions Reduced Denaturing (7% SDS-PAGE)
Loading amount 10 µg
Specification HEK293T
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Dr. Junmo Hwang

Verified customer

Submitted Apr 01 2016

Application Western blot
Sample Rat Cell lysate - other (Rat hepatocytes)
Gel Running Conditions Reduced Denaturing (8% SDS-PAGE)
Loading amount 20 µg
Specification Rat hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 01 2015

Application Immunoprecipitation
Sample Rat Tissue lysate - whole (kidney)
Total protein in input 500 µg
Immuno-precipitation step Protein G
Specification kidney
Username

Gustavo Frindt

Verified customer

Submitted Jul 02 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-15% SDS-PAGE)
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

Submitted Nov 19 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: rt°C
Sample Mouse Cell (nih3t3)
Specification nih3t3
Permeabilization Yes - triton X100 0.5%
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 31 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Sample Rat Cell (Rat hepatocytes)
Specification Rat hepatocytes
Permeabilization No
Fixative Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Oct 28 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Monkey Cell (Kidney)
Permeabilization No
Specification Kidney
Blocking step 3% BSA and 0.5% TX100 as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Dr. Aaron Halpen

Verified customer

Submitted Sep 29 2014

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