Overview

  • Product name
  • Description
    Rabbit polyclonal to SMARCA6
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (Human) - which represents a portion of human Helicase Lymphoid Specific encoded in part by exons 15 and 16.

  • Positive control
    • Nuclear extract from HeLa cells.
  • General notes


    DNA helicases (Hel) play a role in a number of processes involving DNA strand separation, including replication, repair, recombination, and transcription. Jarvis et al. (1996) identified a mouse gene encoding a novel putative helicase. They designated the gene LSH, for lymphoid-specific Hel, because Northern blot analysis revealed that it was expressed specifically in mouse early thymocytes. Geiman et al. (1998) determined that the mouse LSH, or HELLS (helicase lymphoid-specific), gene contains at least 18 exons and spans more than 26.6 kb. Using Northern blot analysis, these authors found that human HELLS is expressed in testis and thymus, two tissues that are highly active in DNA recombination. Geiman and Muegge (2000) demonstrated that LSH is not obligatory for normal lymphoid development but is essential for normal proliferation of peripheral T lymphocytes.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    DNA helicases (Hel) play a role in a number of processes involving DNA strand separation, including replication, repair, recombination, and transcription. Jarvis et al. (1996) identified a mouse gene encoding a novel putative helicase. They designated the gene LSH, for lymphoid-specific Hel, because Northern blot analysis revealed that it was expressed specifically in mouse early thymocytes. Geiman et al. (1998) determined that the mouse LSH, or HELLS (helicase lymphoid-specific), gene contains at least 18 exons and spans more than 26.6 kb. Using Northern blot analysis, these authors found that human HELLS is expressed in testis and thymus, two tissues that are highly active in DNA recombination. Geiman and Muegge (2000) demonstrated that LSH is not obligatory for normal lymphoid development but is essential for normal proliferation of peripheral T lymphocytes.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3851 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 Use at an assay dependent dilution.
WB 1/2000 - 1/10000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).
IP Use a concentration of 2 - 4 µg/ml.

Target

  • Function
    Plays an essential role in normal development and survival. Involved in regulation of the expansion or survival of lymphoid cells. Required for de novo or maintenance DNA methylation. May control silencing of the imprinted CDKN1C gene through DNA methylation. May play a role in formation and organization of heterochromatin, implying a functional role in the regulation of transcription and mitosis.
  • Tissue specificity
    Highly expressed in proliferative tissues such as adult thymus and testis, and expressed at lower levels in uterus, small intestine, colon, and peripheral blood mononuclear cells. Also expressed in neoplastic cell lines including those derived from myeloid and lymphoid leukemias.
  • Sequence similarities
    Belongs to the SNF2/RAD54 helicase family.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
  • Cellular localization
    Nucleus. Closely associated with pericentric heterochromatin.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ10339 antibody
    • Helicase lymphoid specific antibody
    • Hells antibody
    • HELLS_HUMAN antibody
    • LSH antibody
    • Lymphoid specific helicase antibody
    • Lymphoid-specific helicase antibody
    • Nbla10143 antibody
    • PASG antibody
    • Proliferation associated SNF2 like protein antibody
    • Proliferation-associated SNF2-like protein antibody
    • SWI/SNF2 related matrix associated actin dependent regulator of chromatin subfamily A member 6 antibody
    • SWI/SNF2-related matrix-associated actin-dependent regulator of chromatin subfamily A member 6 antibody
    see all

Images

  • Detection of Human HELLS by Western Blot of Immunoprecipitate using Affinity Purified Rabbit anti-HELLS (ab3851).

    Sample: Nuclear Extract (1mg) from HeLa cells

    Antibody: Affinity purified Rabbit anti-HELLS (ab3851)

    IP - 2µg. WB - 0.5µg/ml

    Detection: ECL, 30 seconds

     

  • Anti-SMARCA6 antibody (ab3851) at 1 µg/ml + Human thymus tissue lysate - total protein (ab30146) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 97 kDa
    Observed band size: 97 kDa
    Additional bands at: 102 kDa, 120 kDa, 47 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds

References

This product has been referenced in:
  • Jenness C  et al. HELLS and CDCA7 comprise a bipartite nucleosome remodeling complex defective in ICF syndrome. Proc Natl Acad Sci U S A 115:E876-E885 (2018). Read more (PubMed: 29339483) »
See 1 Publication for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for confirming these details. I am sorry to hear that these antibodies did not work in ICC/IF.
As requested, I have issued a free of charge replacement of ab3851, ab52060, and ab112511. These will be shipped out tonight for delivery tomorrow morning. It looks like that that original order was shipped on a Saturday for Monday's delivery. If you would prefer for us not to send orders on the weekend in the future, please let us know and we can make a note on your account.
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.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7 and HEK)
Loading amount
30 µg
Specification
MCF7 and HEK
Gel Running Conditions
Reduced Denaturing (8-16%)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Oocytes)
Loading amount
100 cells
Specification
Oocytes
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Spermatogenic cells)
Specification
Spermatogenic cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.15% Triton X 100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 29 2007

Question

Here is a complaint about ab3851/Anti-SMARCA6 with lot No 74811. Following is the detail. 1. Please describe the problem at first(when received the antibody) can get result with high background; but when stored it for 2 month, can’t get any bands. 2. On what material are you testing the antibody in WB? Species: human diploid fibroblast(young,middle,old) Cell extract or Nuclear extract: total Cell extract 3. The lysate How much protein was loaded: 120ug What lysis buffer was used: 50mM Tris-HCl,pH7.6?250mM NaCl? 5mM EDTA50mM NaF with protease inhibitors before using What protease inhibitors were used: 1% 100mM PMSF, appropriate leupeptin and aprotinin,10% NP-40 What loading buffer was used: 2×SDS loading buffer add beta-Me before using Did you heat the samples: temperature and time: 90-100C for 3-5mins 4. Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: Reducing Gel Gel percentage : 4% upper SDS-PAGE and 7.5% down SDS-PAGE Transfer conditions: Use Bio-Rad Trans-Blot Cell; Transfer is done at 250mA for 1.5hours on ice.; Transfer buffer----------Glycine 14.4g; methanol 100-200ml; Tris-base 3.03g; ddH2O up to 1000ml,keep in 4C 5. Blocking conditions Buffer: TBS-T(0.02% Tween-20) Blocking agent: milk, BSA, serum, what percentage: 5% milk Incubation time: 2hours Incubation temperature: RT 6. Primary Antibody Specification (in which species was it raised against): rabbit At what dilution(s) have you tested this antibody: 1:2000, 1:2500, 1:5000, 1:8000 What dilution buffer was used: TBS-T containing 1% no-fat milk Incubation time: overnight Incubation temperature: 4C What washing steps were done: 1×TBS-T for first time, in RT for 5min,then blocking buffer for 15min followed by 4 times wash with 1×TBS-T, each time 5min. All the times with shaking. 7. Secondary Antibody Specification (in which species was it raised against)? Goat anti rabbit IgG, HRP At what dilution(s) have you tested this antibody: 1:2000-1:5000 Incubation time: 4C for 1hours Wash steps: 1×TBS-T for first time, in RT for 5min,then blocking buffer for 15min followed by 4 times wash with 1×TBS-T, each time 5min. All the times with shaking Do you know whether the problems you are experiencing come from the secondary?: not from the secondary antibody for it worked well with other primary antibody 8. Detection method ECL 9. Background bands Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) yes 11. Did you apply positive and negative controls along with the samples? With primary antibody against b-actin, can get positive result. Using non-relative protein as negative control, get no bands Attchement is the result of before, with high background: from left to right: extract from Young 2BS,Middle 2BS, Old 2BS, Non-realtive protein

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Answer

I'm sorry to hear you are having a problem with ab3851. Thank you for taking the time to answer our questions and sending us an image of your blot, it is very helpful. I suspect the high background you had in the first blot is due to the presence of milk in the blocking step as well as in the dilution and washing buffers. I would recommend trying 5%BSA blocking and incubating the antibodies in TBST only. I would recommend testing a positive control of nuclear extract of Hela cell lysate to make sure levels of SMARCA6 in your samples are not below detection levels and to use ECL+ as this is more sensitive. The fact that the antibody did not work the second time after 2 months is very concerning, was the antibody stored at 4C and did it stay at room temperature for any significant amount of time? If you still have problems with these recommendations and with the positive control I can certainly offer you a replacement vial if you have purchased the antibody in the last 90 days,

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