AB3200
Anti-LIM-1 Antibody
Chemicon®, from rabbit
Synonim(y):
Anty-LIM-1, Anty-LIM1
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About This Item
Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Polecane produkty
pochodzenie biologiczne
rabbit
Poziom jakości
forma przeciwciała
purified immunoglobulin
rodzaj przeciwciała
primary antibodies
klon
polyclonal
reaktywność gatunkowa
human, frog, fish, mouse
producent / nazwa handlowa
Chemicon®
metody
immunocytochemistry: suitable
immunofluorescence: suitable
immunohistochemistry: suitable (paraffin)
immunoprecipitation (IP): suitable
western blot: suitable
numer dostępu NCBI
numer dostępu UniProt
Warunki transportu
wet ice
docelowa modyfikacja potranslacyjna
unmodified
informacje o genach
human ... LHX1(3975)
Specyficzność
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.
Immunogen
C-terminal portion of frog LIM-1 protein.
Zastosowanie
Anti-LIM-1 Antibody detects level of LIM-1 & has been published & validated for use in IF, IP, WB, IC, IH(P).
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
Immunofluorescence 1:100
Optimal working dilutions must be determined by the end user.
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
Immunofluorescence 1:100
Optimal working dilutions must be determined by the end user.
Research Category
Neuroscience
Neuroscience
Research Sub Category
Developmental Neuroscience
Neuronal & Glial Markers
Developmental Neuroscience
Neuronal & Glial Markers
Postać fizyczna
Format: Purified
Purified immunoglobulin
Przechowywanie i stabilność
Maintain at 2-8°C in undiluted aliquots for up to 6 months.
Informacje prawne
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Oświadczenie o zrzeczeniu się odpowiedzialności
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.
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Kod klasy składowania
10 - Combustible liquids
Klasa zagrożenia wodnego (WGK)
WGK 2
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
Certyfikaty analizy (CoA)
Poszukaj Certyfikaty analizy (CoA), wpisując numer partii/serii produktów. Numery serii i partii można znaleźć na etykiecie produktu po słowach „seria” lub „partia”.
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Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.
Identification of self-replicating multipotent progenitors in the embryonic nervous system by high Notch activity and Hes5 expression.
Onur Basak,Verdon Taylor
The European Journal of Neuroscience null
Dario Acampora et al.
Mechanisms of development, 126(10), 882-897 (2009-07-21)
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Neurobiology of Aging null
A A Karavanov et al.
The International journal of developmental biology, 40(2), 453-461 (1996-04-01)
Polyclonal antibodies to Xlim-1 homeodomain protein of Xenopus laevis were used to study the developmental expression pattern of this protein in Xenopus, rat and mouse. Western blotting of embryo extracts injected with different Xlim-1 constructs confirmed the specificity of the
The sonic hedgehog signaling pathway is reactivated in human renal cell carcinoma and plays orchestral role in tumor growth.
Dormoy, V; Danilin, S; Lindner, V; Thomas, L; Rothhut, S; Coquard, C; Helwig, JJ; Jacqmin et al.
Molecular Cancer null
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