Overview

  • Product name
    Anti-SMN/Gemin 1 antibody [2B1]
    See all SMN/Gemin 1 primary antibodies
  • Description
    Mouse monoclonal [2B1] to SMN/Gemin 1
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, ICC/IF, ELISA, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human, Xenopus laevis
  • Immunogen

    Recombinant full length protein corresponding to Human SMN/Gemin 1.

Properties

Applications

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.

Application Abreviews Notes
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.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    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.
  • Tissue specificity
    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).
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the SMN family.
    Contains 1 Tudor domain.
  • Cellular localization
    Cytoplasm. Nucleus > gem. Localized in subnuclear structures next to coiled bodies, called Gemini of Cajal bodies.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • 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.

  • 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.

  • All lanes : Anti-SMN/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
    All lanes : 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


    Gemin 1contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • 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
  • 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).
  • Anti-SMN/Gemin 1 antibody [2B1] (ab5831) at 1/100 dilution + Xenopus laevis st II-III oocytes whole cell lysate at 20 µg

    Secondary
    Goat Anti-Mouse IgG Fc (HRP) (ab97265) at 1/25000 dilution

    Performed under reducing conditions.

    Predicted band size: 35 kDa
    Observed band size: 31 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

References

This product has been referenced in:
  • Chen Z  et al. LYTAK1 attenuates proliferation of retinal pigment epithelial cells through TGF-ß-mediated epithelial-mesenchymal transition via the ERK/AKT signaling pathway. Exp Ther Med 14:4951-4957 (2017). Read more (PubMed: 29201199) »
  • 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) »
See all 3 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Cell lysate - whole cell (st II-III oocytes)
Gel Running Conditions
Reduced Denaturing (4-20% precast)
Loading amount
20 cells
Specification
st II-III oocytes
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

Ms. Samantha Jeschonek

Verified customer

Submitted Feb 03 2017

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

Abcam user community

Verified customer

Submitted Jul 24 2012

Answer

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 has caused. As requested, I have issued a free of charge replacement with the order number xxx for 1 vial of ab5831 with another lot.

Please let me know which other Gemin1-antibody you would like to receive. We have one that was tested in ICC and with human samples: ab108531 (https://www.abcam.com/Gemin-1-antibody-EPR4429-ab108531.html). This one is currently backordered for 1-2 weeks. Please let me know if I shall arrange for shipping this antibody or if you prefer a different one.


To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

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 antibody?
3) Which blocking reagent was used?
4) Is the secondary antibody working well with other primaries?
5) Did you try out both vials? If not, I'd suggest trying the second vial as well as the issue could be just vial -related.
6) Were there any changes donein your sample preparation or protocol when using the old lot and the newlot?

What kind of replacement were you looking for? We have 1 vial of a different lot in stock currently which I can send you. Alternatively, a different Gemin1 antibody can be sent free of charge.
Please let me what you would prefer. Thank you so much!

I look forward to hear back from you and assist you further.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

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 difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Read More

Answer

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 little more about the situation, so we can better assist you?

1) What lot(s) have you used in the past, and what lot is not working now?
2) What species are the spinal cord sections from?
3) Is there any staining at all, or is the staining pattern incorrect?
4) Are you using the same tissue as you did with the previous vial(s)? If so, how has the tissue been stored?
5) What dilutions or concentrations have you tried? Did you leave the antibody on the tissue overnight or for a shorter incubation time?
6) What kind of antigen retrieval was tried?
7) What blocking solution is used?

I look forward to hearing from you and resolving this issue promptly. Please let me know if you have any further questions or if there is anything else that we can do for you.

Read More
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

Abcam user community

Verified customer

Submitted Apr 26 2011

Question
Answer

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 experiment to determine an optimal concentration: try a few dilutions of 1:500, 1:1000, 1:2000. Then optimise the dilution depending on those results. Good luck with your research,

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

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