Anti-Gemin 1抗体[2B1] (ab5831)

概述

  • 产品名称Anti-Gemin 1抗体[2B1]
    参阅全部 Gemin 1 一抗
  • 描述
    小鼠单克隆抗体[2B1] to Gemin 1
  • 经测试应用适用于: IHC-P, ICC/IF, ELISA, WB, IP, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Human, Xenopus laevis
  • 免疫原

    Recombinant full length His 6 tagged SMN protein (Human).

性能

应用

Our Abpromise guarantee covers the use of ab5831 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 35 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

靶标

  • 功能The SMN complex plays an essential role in spliceosomal snRNP assembly in the cytoplasm and is required for pre-mRNA splicing in the nucleus. It may also play a role in the metabolism of snoRNPs.
  • 组织特异性Expressed in a wide variety of tissues. Expressed at high levels in brain, kidney and liver, moderate levels in skeletal and cardiac muscle, and low levels in fibroblasts and lymphocytes. Also seen at high levels in spinal cord. Present in osteoclasts and mononuclear cells (at protein level).
  • 疾病相关Defects in SMN1 are the cause of spinal muscular atrophy autosomal recessive type 1 (SMA1) [MIM:253300]. Spinal muscular atrophy refers to a group of neuromuscular disorders characterized by degeneration of the anterior horn cells of the spinal cord, leading to symmetrical muscle weakness and atrophy. Autosomal recessive forms are classified according to the age of onset, the maximum muscular activity achieved, and survivorship. The severity of the disease is mainly determined by the copy number of SMN2, a copy gene which predominantly produces exon 7-skipped transcripts and only low amount of full-length transcripts that encode for a protein identical to SMN1. Only about 4% of SMA patients bear one SMN1 copy with an intragenic mutation. SMA1 is a severe form, with onset before 6 months of age. SMA1 patients never achieve the ability to sit.
    Defects in SMN1 are the cause of spinal muscular atrophy autosomal recessive type 2 (SMA2) [MIM:253550]. SMA2 is an autosomal recessive spinal muscular atrophy of intermediate severity, with onset between 6 and 18 months. Patients do not reach the motor milestone of standing, and survive into adulthood.
    Defects in SMN1 are the cause of spinal muscular atrophy autosomal recessive type 3 (SMA3) [MIM:253400]. SMA3 is an autosomal recessive spinal muscular atrophy with onset after 18 months. SMA3 patients develop ability to stand and walk and survive into adulthood.
    Defects in SMN1 are the cause of spinal muscular atrophy autosomal recessive type 4 (SMA4) [MIM:271150]. SMA4 is an autosomal recessive spinal muscular atrophy characterized by symmetric proximal muscle weakness with onset in adulthood and slow disease progression. SMA4 patients can stand and walk.
  • 序列相似性Belongs to the SMN family.
    Contains 1 Tudor domain.
  • 细胞定位Cytoplasm. Nucleus > gem. Localized in subnuclear structures next to coiled bodies, called Gemini of Cajal bodies.
  • Information by UniProt
  • 数据库链接
  • 别名
    • BCD541 antibody
    • Component of gems 1 antibody
    • Gemin 1 antibody
    • Gemin-1 antibody
    • OTTHUMP00000125198 antibody
    • OTTHUMP00000223567 antibody
    • OTTHUMP00000223568 antibody
    • OTTHUMP00000224066 antibody
    • OTTHUMP00000226924 antibody
    • SMA 1 antibody
    • SMA 2 antibody
    • SMA 3 antibody
    • SMA 4 antibody
    • SMA antibody
    • SMA@ antibody
    • SMA1 antibody
    • SMA2 antibody
    • SMA3 antibody
    • SMA4 antibody
    • SMN antibody
    • SMN_HUMAN antibody
    • SMN1 antibody
    • SMN2 antibody
    • SMNT antibody
    • Survival motor neuron protein antibody
    • Survival of motor neuron 1, telomeric antibody
    • T-BCD541 antibody
    see all

Anti-Gemin 1 antibody [2B1] 图像

  • All lanes : Anti-Gemin 1 antibody [2B1] (ab5831) at 1 µg/ml

    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 35 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 45 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • IHC image of ab5831 staining in human normal cervical carcinoma 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 ab5831, 5µg/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.

  • Overlay histogram showing HepG2 cells stained with ab5831 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5831, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ab5831 staining Gemin 1 in human HeLa cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol and then blocked using 0.2% fish scale gelatin for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/300 for 20 minutes at 25°C. The secondary antibody used was a donkey anti-mouse IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. Counterstained with DAPI (blue).
    Gemin 1 is clearly visible in the cytoplasm and also as small dots in the nucleus (cajal bodies).
  • Gemin 1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Gemin 1 (ab5831) 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,Hela 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 ab5831.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 32kDa: Gemin 1
  • ICC/IF image of ab5831 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 (ab5831, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

Anti-Gemin 1 antibody [2B1] (ab5831)参考文献

This product has been referenced in:
  • Schertzer M  et al. Human regulator of telomere elongation helicase 1 (RTEL1) is required for the nuclear and cytoplasmic trafficking of pre-U2 RNA. Nucleic Acids Res 43:1834-47 (2015). Human . Read more (PubMed: 25628358) »
  • Ishihara T  et al. Decreased number of Gemini of coiled bodies and U12 snRNA level in amyotrophic lateral sclerosis. Hum Mol Genet 22:4136-47 (2013). Read more (PubMed: 23740936) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (ES cells)
Specification ES cells
Fixative Methanol
Permeabilization No
Blocking step Fish scale gelatin in PBS as blocking agent for 20 minute(s) · Concentration: 0.2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 24 2012

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this ...

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Thank you for contacting us.

Could you please send me a few more experimental details for our records:

1) What samples and species was used?
2) What is the antibody dilution and incubation time employed for the primary antibo...

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According to our records, ab5831 was proving difficult to use and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the di...

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Thank you for contacting us with your question and for letting us know about the trouble with this new vial of ab5831.

I have looked through our records, and we don't have any other reports of this antibody not working. Could you tell me a li...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela cells)
Specification Hela cells
Fixative Methanol
Permeabilization No
Blocking step Fish scale gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Apr 26 2011

Thank you for your enquiry. The datasheet does not recommend a specific dilution as it may vary between preparations/tissues and also depending on the amount of gemin 1 in your sample and the detection system you use. I would recommend a preliminary e...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"