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, Marmoset (common)
    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
    • E14tG2a (Mouse embryonic stem cell line) Whole Cell Lysate. IHC-P: FFPE Human Testis Normal

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 1 - 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

Anti-Sall4 antibody - ChIP Grade 图像

  • 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

  • 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)

  • 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).

Anti-Sall4 antibody - ChIP Grade (ab29112)参考文献

This product has been referenced in:
  • Yu-Ching Lin Z  et al. Gene expression ontogeny of spermatogenesis in the marmoset uncovers primate characteristics during testicular development. Dev Biol N/A:N/A (2015). IHC-P ; Marmoset (common) . Read more (PubMed: 25624265) »
  • Zheng Y  et al. Quantitative detection of human spermatogonia for optimization of spermatogonial stem cell culture. Hum Reprod 29:2497-511 (2014). Read more (PubMed: 25267789) »

See all 16 Publications for this product

Product Wall

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

Thanks so much for your reply. My colleague is out of the office today, but I'll be happy to assist you with this.

I've issued credit note number *** for your original order of ab29112. Since this order was placed with a credit card, the purc...

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Thank you for your reply.

I am sorry to hear that the protocol suggestions did not improve the staining. I have found your original order and would be happy to offer you a free of charge replacement antibody or refund/credit note.

P...

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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

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for...

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