Alpha-Tubulin Antibody, Unconjugated, Rabbit, Polyclonal Preis auf Anfrage
Biozol Catalog Number:
BYT-ORB345510
Supplier Catalog Number:
orb345510
Alternative Catalog Number:
BYT-ORB345510-200
Manufacturer:
Biorbyt
Host:
Rabbit
Category:
Antikörper
Application:
ELISA, IF, IHC, WB
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-Tubulin Loading Control Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to the C-Terminal region near amino acids 425-451 of Human alpha Tubulin.
Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None, Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Purity:
Anti-Tubulin Loading Control Antibody is directed against human alpha Tubulin protein. The Loading Control Antibody was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest that this antibody would react with alpha Tubulin from a wide range of organisms, including avian, mammalian aquatic, parasitic and alga sources based on 100% homology for the immunogen sequence. Cross reactivity will occur with all isoforms of alpha tubulin. Such broad reactivity makes this antibody useful as an excellent loading control.
Application Notes: Anti-Tubulin Antibody has been tested for use in ELISA, immunofluorescence, and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band at ~50 kDa in size corresponding to alpha tubulin by western blotting in most cell lysates or extracts
ELISA results of purified Rabbit anti-alpha-Tubulin Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1 µg of conjugate. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n orb347654) and TMB ELISA Peroxidase Substrate (p/n orb348651).
Immunofluorescence microscopy of Rabbit Anti-alpha-Tubulin antibody using HeLa cells fixed with PFA. Anti-alpha-Tubulin Antibody was used at 1 µg/ml, O/N at 4°C. Secondary antibody: Anti-RABBIT IgG DyLight(TM) 488 Conjugated Preadsorbed at 2 ug/ml for 1 h at RT. Localization: TUBA1B is the major constituent of microtubules in the cytoplasm. Staining: Tubulin as green fluorescent signal with DAPI (blue) nuclear counterstain.
miR-155 and FOXP3 down regulate endogenous ZEB2 in human breast cancer cells resulting in altered levels of EMT markers Vimentin and E-cadherin(A) Relative abundance of ZEB2 and ZEB1 protein in WT, GFP or FOXP3 overexpressing BT549 cells transfected with miR-155 or miR-control. Relative abundance of protein was determined by quantitation of the abundance of ZEB2 or ZEB1 proteins normalised to reference protein alpha-Tubulin by western blot analysis. Quantitation of bands was carried out using Image J software. Mean + SD plotted. Students t test ***P < 0.001. ZEB1 protein expression as above. n = 3 experiments. (B) ZEB2 and ZEB1 protein in WT, GFP or FOXP3 overexpressing BT549 cells transfected with miR-155 or miR-control by western blot. Representative western blot shown. (C) Relative abundance of Vimentin and E-cadherin protein in WT, GFP or FOXP3 overexpressing BT549 cells transfected with miR-155 or miR-control. Relative abundance of protein was determined by quantitating the abundance of E-cadherin or Vimentin proteins and normalising to reference protein beta-Actin by western blot analysis. Quantitation of bands was carried out using Image J software. Mean + SD plotted. Students t test ***P < 0.001, **P < 0.01. n = 3 experiments. (D) Vimentin and E-cadherin protein in WT, GFP or FOXP3 overexpressing BT549 cells transfected with miR-155 or miR-control analysed by western blot. Representative western blot shown.
Western Blot of Rabbit Anti-Alpha Tubulin Antibody. Lane 1: whole cell lysates from mouse brain (p/n orb348715). Lane 2: rat brain (p/n orb348727). Lane 3: A431 cells (p/n orb348665). Lane 4: Jurkat cells (p/n orb348674). Lane 5: HeLa cells (p/n orb348668). Load: 35 µg per lane. Primary antibody: Alpha Tubulin antibody at 1:1200 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO (p/n orb348624) overnight at 4C. Predicted/Observed size: ~50 kDa corresponding to alpha tubulin (arrowhead). Other band(s): none.
Western Blot of Rabbit anti-alpha-Tubulin antibody. Lane 1: HeLa WCL (p/n orb348668). Lane 2: NIH/3T3 WCL (p/n orb348714). Load: 10 µg per lane. Primary antibody: alpha-Tubulin antibody at 1:1000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n orb348637) for 30 min at RT. Predicted/Observed size: 50 kDa, 50 kDa for alpha-Tubulin. Other band(s): N/A.
Western Blot of Rabbit anti-Alpha-Tubulin antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate (p/n orb348669). Lane 2: HeLa Lysate (p/n orb348668). Lane 3: MCF-7 Lysate (p/n orb348664). Lane 4: Jurkat Lysate. Lane 5: A431 Lysate (p/n orb348665). Lane 6: LNCaP Lysate (p/n orb348694). Lane 7: A-172 Lysate (p/n orb348708). Lane 8: NIH/3T3 Lysate (p/n orb348714). Load: 35 µg per lane. Primary antibody: Alpha-Tubulin antibody at 1:2000 for overnight at 4C. S
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