Anti-EB1 antibody produced in rabbit
SIGMA/E3406 - ~1.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Synonym: Anti-APC binding protein EB1; Anti-End-Binding-1; Anti-
MDL Number: MFCD01633308
Product Type: Chemical
Immunoblotting
RIPA cell lysates of various cell lines were separated on SDS-PAGE and blotted with Anti-EB1 (Cat. No. E3406). The antibody was developed with Goat Anti-Rabbit IgG, Peroxidase conjugate (Cat. No. A0545) and a chemiluminescent substrate. Antibody dilution 1:1,000.
Lanes
1. HeLa
2. NIH3T3
3. Rat1
4. A431
5. SW48
6. HCT116
7. HEK293
8. HeLa. Inhibition of antibody binding with EB1 immunizing peptide (human, 252-268) (10 μg/mL)
RIPA cell lysates of various cell lines were separated on SDS-PAGE and blotted with Anti-EB1 (Cat. No. E3406). The antibody was developed with Goat Anti-Rabbit IgG, Peroxidase conjugate (Cat. No. A0545) and a chemiluminescent substrate. Antibody dilution 1:1,000.
Lanes
1. HeLa
2. NIH3T3
3. Rat1
4. A431
5. SW48
6. HCT116
7. HEK293
8. HeLa. Inhibition of antibody binding with EB1 immunizing peptide (human, 252-268) (10 μg/mL)
Immunofluorescence
Staining of the “plus-end tracking protein” (+TIP) structures in Rat1 fibroblasts. Rat fibroblast Rat-1 cells were grown to semi-confluency, fixed with acetone/methanol and stained with Anti-EB1 (Cat. No. E3406). The antibody was developed with Goat Anti-Rabbit IgG, Cy3 conjugate. Antibody dilution 1:1,000.
Staining of the “plus-end tracking protein” (+TIP) structures in Rat1 fibroblasts. Rat fibroblast Rat-1 cells were grown to semi-confluency, fixed with acetone/methanol and stained with Anti-EB1 (Cat. No. E3406). The antibody was developed with Goat Anti-Rabbit IgG, Cy3 conjugate. Antibody dilution 1:1,000.
Immunoprecipitation
RIPA lysates of HeLa cells were immunoprecipitated with Anti-EB1 (Cat. No. E3406), separated on SDS-PAGE and blotted with Anti-EB1 (Cat. No. E3406) at a 1:1,000 dilution. The antibody was developed with Goat Anti-Rabbit IgG, Peroxidase (Cat. No. A0545) and a chemiluminescent substrate. *50kDa band represents Rb IgG heavy chain. (Lane 1) No lysate, no Anti-EB1 (Negative control); (Lane 2) No lysate, + Anti-EB1 (15 μg) (Negative control); (Lane 3) Cell lysate (100 μg) + Anti-EB1 (15 μg); (Lane 4) Cell lysate (25 μg) + Anti-EB1 (15 μg).
RIPA lysates of HeLa cells were immunoprecipitated with Anti-EB1 (Cat. No. E3406), separated on SDS-PAGE and blotted with Anti-EB1 (Cat. No. E3406) at a 1:1,000 dilution. The antibody was developed with Goat Anti-Rabbit IgG, Peroxidase (Cat. No. A0545) and a chemiluminescent substrate. *50kDa band represents Rb IgG heavy chain. (Lane 1) No lysate, no Anti-EB1 (Negative control); (Lane 2) No lysate, + Anti-EB1 (15 μg) (Negative control); (Lane 3) Cell lysate (100 μg) + Anti-EB1 (15 μg); (Lane 4) Cell lysate (25 μg) + Anti-EB1 (15 μg).
Immunofluorescence Anti-EB1: Cat. No. E3406: Immunofluorescence of HUVEC cells using Anti-EB1 (red) at a 1:200 dilution, taken at 40× magnification and nuclear staining with Hoescht 33342 (blue). Yale HTCB IF procedure used.
Immunofluorescence Graph Anti-EB1: Cat. No. E3406: Intensity analysis of Anti-EB1 staining. Cytoplasmic and nuclear intensity values were obtained at 8 different antibody dilutions (1:50 to 1:6,400) as compared with the negative control. Yale HTCB IF procedure used.
| antibody form | affinity isolated antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | polyclonal |
| concentration | ~1.5 mg/mL |
| conjugate | unconjugated |
| form | buffered aqueous solution |
| mol wt | antigen 35 kDa |
| Quality Level | 200  |
| shipped in | dry ice |
| species reactivity | rat, mouse, bovine, pig, canine, human |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunoprecipitation (IP): 10-20 μg using cell lysate of the human epitheloid carcinoma HeLa cell line |
| indirect immunofluorescence: 1-2 μg/mL using rat fibroblast Rat1 cells | |
| indirect immunofluorescence: suitable | |
| western blot: 0.5-1 μg/mL using cell lysate of the rat fibroblast cell line Rat1 or Rat 2 | |
| UniProt accession no. | Q15691  |
| Application: | Anti-EB1 antibody produced in rabbit has been used in • immunofluorescence • western blotting • immunocytochemistry • immunoprecipitation |
| Biochem/physiol Actions: | End-Binding-1 (EB1) associates with components of the dynactin complex and cytoplasmic dynein intermediate chain. |
| Biochem/physiol Actions: | The C-terminal domain of End-binding proteins (EBs) is important for its autonomous microtubule tip tracking, regulation of microtubule dynamics, chromosome segregation and recruitment of numerous partners to microtubule ends. EB1 proteins bind to microtubule ends and act with the adenomatous polyposis coli (APC) tumor suppressor to regulate the microtubule filament system. It is a potential oncogene and plays an important role during early event in colon carcinogenesis, colorectal cancer (CRC). EB1 acts as a potential biomarker and therapeutic target in CRC. |
| Disclaimer: | 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. |
| General description: | Microtubule-associated protein RP/EB family member 1 is a protein encoded by the MAPRE1 gene in humans. EB1 (End-Binding-1) proteins belong to a distinct family of accessory proteins, termed “plus-end tracking proteins” or +TIPs, that bind to the plus-ends of microtubules. All EB1 family proteins have a highly conserved N-terminal region that contains a calponin homology (CH) domain and an EB1-like C-terminal motif, which is unique to EB1 family proteins. |
| Immunogen: | synthetic peptide corresponding to amino acids 252-268 located at the C-terminus of human EB1, conjugated to KLH. This sequence is identical in several species (mouse, rat, dog, bovine, and porcine EB1), highly conserved (single amino acid substitution) in chicken EB1, and has limited homology to human EB2 (35% identity) and human EB3 (53% identity). |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |