Anti-Sall4抗体- ChIP Grade (ab29112)

概述

  • 产品名称
    Anti-Sall4抗体- ChIP Grade
    参阅全部 Sall4 一抗
  • 描述
    兔多克隆抗体to Sall4 - ChIP Grade
  • 经测试应用
    适用于: ChIP/Chip, IHC-Fr, WB, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Human, Macaque monkey, Common marmoset
    预测可用于: Chicken, Xenopus laevis
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1000 to the C-terminus of Mouse Sall4.

  • 阳性对照
    • WB: E14tG2a (Mouse embryonic stem cell line) whole cell lysate. IHC-P: Human normal testis tissue. ICC/IF: Mouse embryonic stem cells.

性能

应用

Our Abpromise guarantee covers the use of ab29112 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
ChIP/Chip Use at an assay dependent concentration.
IHC-Fr 1/1000.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 85, 140 kDa (predicted molecular weight: 66, 113 kDa).
IHC-P Use a concentration of 0.5 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.

靶标

  • 功能
    Probable transcription factor.
  • 组织特异性
    Expressed in testis.
  • 疾病相关
    Defects in SALL4 are the cause of Duane-radial ray syndrome (DRRS) [MIM:607323]; also known as Okihiro syndrome. DRRS is a disorder characterized by the association of forearm malformations with Duane retraction syndrome.
    Defects in SALL4 are the cause of oculootoradial syndrome (OORS) [MIM:147750]. Oculootoradial syndrome is an autosomal dominant condition characterized by upper limbs anomalies (radial ray defects, carpal bones fusion), extraocular motor disturbances, congenital bilateral non-progressive mixed hearing loss. Other less consistent malformations include heart involvement, mild thrombocytopenia and leukocytosis (before age 50), shoulder girdle hypoplasia, imperforate anus, kidney malrotation or rectovaginal fistula. The IVIC syndrome is an allelic disorder of Duane-radial ray syndrome (DRRS) with a similar phenotype.
  • 序列相似性
    Belongs to the sal C2H2-type zinc-finger protein family.
    Contains 7 C2H2-type zinc fingers.
  • 细胞定位
    Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • AA407717 antibody
    • AL022809 antibody
    • AW536104 antibody
    • C330011P20Rik antibody
    • C78083 antibody
    • C78563 antibody
    • dJ1112F19.1 antibody
    • DRRS antibody
    • HSAL4 antibody
    • Sal like 4 (Drosophila) antibody
    • Sal like 4 antibody
    • Sal like Protein 4 antibody
    • Sal-like protein 4 antibody
    • Sall4 antibody
    • SALL4_HUMAN antibody
    • Spalt like transcription factor 4 antibody
    • Tex20 antibody
    • Zinc finger protein 797 antibody
    • Zinc finger protein SALL4 antibody
    • ZNF797 antibody
    see all

图片

  • IHC image of ab29112 staining in human testis 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 ab29112, 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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

  • ICC/IF image of ab29112 stained human embryonic stem cells. The cells were fixed in Paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 10% Goat serum, 0.1% BSA in PBS for 1 hour at RT, before incubation with ab29112 at a 1/100 dilution for 12 hours at 4°C. The secondary used was an Alexa Fluor® 488 conjugated goat anti-rabbit polyclonal, at 1/200 dilution.

    See Abreview

  • All lanes : Anti-Sall4 antibody - ChIP Grade (ab29112) at 1 µg/ml

    Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 66, 113 kDa
    Observed band size : 80,140 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa (possible non-specific binding).

    Exposure time : 8 minutes

    MEF1 whole cell lysate was included as a negative control. 

     

    Secondary antibody - goat anti-rabbit HRP (ab97051)

  • IHC-P image of Sall4 staining with ab29112 on tissue sections from a 3 year old adult marmoset testis. The sections were subjected to heat-mediated antigen retrieval using Dako antigen retrieval solution. The sections were then blocked with 5% milk for 30 minutes at 25°C, before incubation with ab29112 (1/100 dilution) for 20 hours at 4°C. The secondary was an Alexa Fluor® 555 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

    See Abreview

  • Anti-Sall4 antibody - ChIP Grade (ab29112) at 1 µg + E14tG2a (Mouse embryonic stem cell line) Whole Cell Lysate at 20 µg

    Secondary
    IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

    Performed under reducing conditions.

    Predicted band size : 66, 113 kDa
    Observed band size : 85,140 kDa (why is the actual band size different from the predicted?)
    This antibody detects two bands which we believe correspond to the two isoforms of Sall4 outlined in Ma et al, 2006 (PMID: 16763212). This paper describes human isoforms running at 165 kDa and 95 kDa. In mouse embryonic stem cells we see both isoforms running at slightly lower molecular weights. We believe that these bands represent Sall4a (Swissprot: Q8BX22) and Sall4b (Swissprot: Q6S7E9) which are predicted to be 113 kDa and 66 kDa respectively.
  • ICC/IF image of ab29112 stained mouse embryonic stem 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 (ab29112, 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).

文献

This product has been referenced in:
  • Zhou W  et al. MicroRNA-98 acts as a tumor suppressor in hepatocellular carcinoma via targeting SALL4. Oncotarget 7:74059-74073 (2016). WB ; Human . Read more (PubMed: 27677076) »
  • Wang F  et al. Leukemic survival factor SALL4 contributes to defective DNA damage repair. Oncogene 35:6087-6095 (2016). IHC-P ; Human . Read more (PubMed: 27132514) »

See all 34 Publications for this product

客户评价及客户问答

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Spermatogonia)
Permeabilization
Yes - sodium citrate-98¯C-12min-5. Permeabilized in 0.2% Triton X-100 in PBS-30 min
Specification
Spermatogonia
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 10 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Mr. Bryan Niedenberger

Verified customer

提交于 Apr 04 2016

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (undifferentiated human iPS cells)
Specification
undifferentiated human iPS cells
Blocking step
5% milk in TBST as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Dec 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Sample
Human Cell (Human Cell - feeder-free human ES cells)
Specification
Human Cell - feeder-free human ES cells
Permeabilization
Yes - see blocking buffer
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (adult testis)
Specification
adult testis
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

煜欽 林

Verified customer

提交于 Jan 02 2013

Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Human Embryonic Stem cells)
Loading amount
70000 cells
Specification
Human Embryonic Stem cells
Gel Running Conditions
Reduced Denaturing (10% SDS)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Dec 12 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (D3 Mouse Embryonic Stem Cell)
Specification
D3 Mouse Embryonic Stem Cell
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 21°C
Username

Joe Segal

Verified customer

提交于 Nov 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Stem Cells)
Loading amount
200000 cells
Specification
Mouse Embryonic Stem Cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Joe Segal

Verified customer

提交于 Oct 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Embryonic Stem Cells)
Specification
Human Embryonic Stem Cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Sep 24 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (adult testis)
Specification
adult testis
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Antigen retrieval solution
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

煜欽 林

Verified customer

提交于 Aug 29 2012

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