重组
RabMAb

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

概述

  • 产品名称
    Anti-GM130抗体[EP892Y] - cis-Golgi Marker
    参阅全部 GM130 一抗
  • 描述
    兔单克隆抗体[EP892Y] to GM130 - cis-Golgi Marker
  • 特异性
    Mouse 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.
  • 经测试应用
    适用于: ICC/IF, IHC-P, IHC-Fr, Flow Cyt, WB, IPmore details
  • 种属反应性
    与反应: Cow, Dog, Human, Monkey, African green monkey
    不与反应: Mouse, Rat
  • 免疫原

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

  • 阳性对照
    • 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 cells, ARPE-19 cells, Bovine brain microvascular endothelial cells, and monkey kidney cells. Flow Cyt: HeLa cells.
  • 常规说明

    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.

    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)

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

性能

应用

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.

应用 Ab评论 说明
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.
Flow Cyt 1/20.
WB 1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 112 kDa).
IP 1/20 - 1/50.

靶标

  • 功能
    Golgi auto-antigen; probably involved in maintaining cis-Golgi structure.
  • 序列相似性
    Belongs to the GOLGA2 family.
  • 结构域
    Extended rod-like protein with coiled-coil domains.
  • 细胞定位
    Golgi apparatus > Golgi stack membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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.

  • 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).

  • 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.

  • 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.

  • 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 liver tissue labelling GM130 with unpurified ab52649 at a dilution of 1/500.

  • 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

  • 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
  • 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

  • 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.

文献

This product has been referenced in:

See all 54 Publications for this product

客户评价及客户问答

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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 Sep 29 2014

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