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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, F, Fg | IP, WB, ICC, IH(P), IF | Rb | Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | AB3200 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-LIM-1 Antibody |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified immunoglobulin |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-LIM-1 Antibody detects level of LIM-1 & has been published & validated for use in IF, IP, WB, IC, IH(P). |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | C-terminal portion of frog LIM-1 protein. |
| Host | Rabbit |
| Specificity | 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. |
| Species Reactivity |
|
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | 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. |
| Gene Symbol |
|
| UniProt Number |
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| UniProt Summary | 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. |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at 2-8°C in undiluted aliquots for up to 6 months. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |