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品种反应性 主要应用 宿主 格式 抗体类型 Fg, H, M, F ICC, WB Rb Purified Polyclonal Antibody 描述 产品目录编号 AB3200 品牌系列 Chemicon®
商名 - Chemicon
描述 Anti-LIM-1 Antibody 产品信息 格式 Purified 演示 Purified immunoglobulin 应用 应用 Anti-LIM-1 Antibody detects level of LIM-1 & has been published & validated for use in IF, IP, WB, IC, IH(P). 主要应用 - Immunocytochemistry
- Western Blotting
应用说明 Immunohistochemistry: 1:200-1:1,000 (Fish and whole mounts 1:500) Recommended fixative MEMFA.
Our photo was produced from immersion fixation of chicken embryos in cold 4% PFA for 15-30 minutes, then sectioning was performed on a cryostat.
Works on paraffin embedded sections when sections are either lightly PFA fixed or fixed with acetone, ethanol or fixative suggested below.
SUGGESTED IMMUNOHISTOCHEMISTRY PROTOCOL FOR AB3200
The best fixative is MEMFA. 10X stock solution for MEMFA: 1 M MOPS, 20 MM EGTA. 10 mM MgSO4, 38% Formaldehyde. Fixation 1 hour, 2 x 15 min methanol. Following this protocol embryos may be stored in methanol at -20°C indefinitely or immediately embedded in paraplast. Best results on paraffin sections 6-10 micron thick.2) Staining following deparaffinization in xylene and a row of alcohol wash two times in water. Block in 2% Boehringer Mannheim reagent in 0.1 M maleic acid, pH 7.5, 150 mM NaCl for one hour at room temperature.3) Dilute the AB3200 in same blocking reagent and incubate overnight at 4°C or for at least 5 hours. Wash three times in PBS, 10 min each.4) Incubate with alkaline phosphatase conjugated secondary antibody (for example Chemicon Catalog Number AP132A. Develop with BCIP/TNBT (Chemicon Catalog Number ES007-100ML).5) For sections always use Digene silanated slides or Superfrost plus from Fisher as some times you may need to boil sections in 6 M urea for 5-6 min. in microwave at 80% power following deparaffinization to increase signal. That is especially useful if tissues were fixed in PFA.6) Sometimes it is necessary to predeplete antibody on hyperfixed embryos to lower background (especially for staining species other than frog and for whole-mounts). Procedure: hyperfix frog or fish embryos in MEMFA for 36-48 hours at RT 30-50 embryos. Wash 2 X in methanol (see above). Apply antibody in final dilution in blocking reagent for one hour on rocking table. Collect super and apply to your embryos or sections. For whole mounts use the following procedure: after fixation, methanol and PBS; block in PBST + serum (PBS + 2 mg/mL BSA + 0.1% Triton X100 + 10% animal serum) one hour room temperature. Add first antibody in PBST + serum and incubate over night at 4°C. Wash in PBST (no serum) 4 times for 2 hours. Add secondary diluted in PBST + serum over night at 4°C. Wash in PBST four times for 2 hours. Develop staining.
Do not use tissues fixed overnight in PFA the antibody will not work.
Immunocytochemistry: 1:500 on P19 cell line, lightly fixed (2% PFA) 5-15 minutes, permeabilized with 0.1% triton X-100 or methanol ( 5' air dry).
Western blot: 1:3,000-1:6,000
Immunoprecipitation: 1:200
Optimal working dilutions must be determined by the end user.生物信息 免疫原品种 C-terminal portion of frog LIM-1 protein. 宿主 Rabbit 特异性 Recognizes LIM-1. This antibody does not appear to cross react with LIM-5 (the closest homologue of LIM-1 in frog) on tissue sections but does cross react with LIM-5 in Western blot and immunoprecipitation. 品种反应性 FrogHumanMouseFish 抗体类型 Polyclonal Antibody Entrez基因编号 - NM_005568.2
Entrez基因汇总 This gene encodes a member of a large protein family which contains the LIM domain, a unique cysteine-rich zinc-binding domain. The encoded protein may function as a transcriptional regulator and be involved in control of differentiation and development of neural and lymphoid cells. A similar protein in mice is an essential regulator of the vertebrate head organizer. 基因符号 - MGC126723
- LHX1
- LIM-1
- hLim-1
- LIM1
- MGC138141
UniProt编号 - P48742
UniProt汇总 FUNCTION: SwissProt: P48742 # Potential transcription factor. May play a role in early mesoderm formation and later in lateral mesoderm differentiation and neurogenesis.
SIZE: 406 amino acids; 44808 Da
SUBUNIT: Interacts with LDB1 via the tandem LIM domains (By similarity).
SUBCELLULAR LOCATION: Nucleus (Probable).
TISSUE SPECIFICITY: Expressed in the brain, thymus, and tonsils. Expressed in samples from patients with chronic myeloid leukemia (CML) and in 58% of acute myeloid leukemia (AML) cell lines.
DOMAIN: SwissProt: P48742 The LIM domains exert a negative regulatory function and disruption of the LIM domains produces an activated form. In addition, two activation domains and a negative regulatory domain exist C-terminally to the homeobox (By similarity).
SIMILARITY: Contains 1 homeobox DNA-binding domain. & Contains 2 LIM zinc-binding domains.产品使用声明 使用声明 - Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
储存和货运信息 存储条件 Maintain at 2-8°C in undiluted aliquots for up to 6 months. 包装信息 数量 100 µg
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下一件merck millipore产品：merck millipore,默克密理博,14-535,Rse Protein, active, 10 µg
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