Anti α-Tubulin  

Immunofluorescence of Rabbit Anti-Alpha Tubulin Antibody. Cell line:？ HeLa？. Primary Antibody: Alpha Tubulin？ (p/n 600-401-880？) at 4.4 μg/mL (1:250) for 1hr at RT？. Secondary Antibody: Goat Anti-Rabbit？ DyLight^TM 488？ (p/n 611-141-121) at 1 μg/mL (1:1000) overnight at 4 °C？. Fixative:？ Ice Cold Methanol？. Permeabilization: Ice Cold Methanol？. Nuclear stain:？ Hoechst 33342？. Expected Localization:？ Cytoplasmic？. Image: A) Alpha Tubulin, B)？ Nuclear Stain？, C) Merge？, D)？ Secondary Only Control？.

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 (p/n 611-741-127) 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.

Western Blot of Rabbit anti-alpha-Tubulin antibody. Lane 1: HeLa WCL (p/n W09-000-364). Lane 2: NIH/3T3 WCL (p/n W10-000-358). Load: 10 μg per lane. Primary antibody: alpha-Tubulin antibody at 1:1,000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n 611-103-122) at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) 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. Lane 1: whole cell lysates from mouse brain (p/n W10-000-T004). Lane 2: rat brain (p/n W12-000-T077). Lane 3: A431 cells (p/n W09-000-361). Lane 4: Jurkat cells (p/n W09-001-370). Lane 5: HeLa cells (p/n W09-000-364). Load: 35 μg per lane. Primary antibody: Alpha Tubulin antibody at 1:1,200 for overnight at 4°C. Secondary antibody: IRDye800^TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO (p/n B501-0500) overnight at 4°C. Predicted/Observed size: ~50 kDa corresponding to alpha tubulin (arrowhead). Other band(s): none.

Western Blot of Rabbit anti-Alpha-Tubulin antibody. Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 1: HEK293 lysate (p/n W09-000-365). Lane 2: HeLa Lysate (p/n W09-000-364). Lane 3: MCF-7 Lysate (p/n W09-000-360). Lane 4: Jurkat Lysate (p/n W09-000-370). Lane 5: A431 Lysate (p/n W09-000-361). Lane 6: LNCaP Lysate (p/n W09-001-GJ9). Lane 7: A-172 Lysate (p/n W09-001-GL5). Lane 8: NIH/3T3 Lysate (p/n W10-000-358). Load: 35 μg per lane. Primary antibody: Alpha-Tubulin antibody at 1:2,000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n 611-103-122) at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS (p/n MB-082) for 30 min at RT. Predicted/Observed size: 50 kDa for Alpha-tubulin.

ELISA results of purified Rabbit anti-alpha-Tubulin Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1μg of conjugate. The starting dilution of antibody was 5μg/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 611-103-122) and TMB ELISA Peroxidase Substrate (p/n TMBE-1000).

Identification of the SIAH2 interaction domain in C/EBPδ. (A) Six different C/EBPδ-C/EBPβ chimeric proteins, as shown in the schematic and the full-length proteins, were cotransfected with Flag-SIAH2 in MCF-10A cells. Immunoprecipitates with anti-Flag antibody and input were analyzed with a pan-C/EBP and anti-Flag antibody for SIAH2. Cells had been treated with MG132 to stabilize C/EBPδ. TA, transactivation domain; BR, basic region; LZ, leucine zipper. (B) Schematic of the sequence of human C/EBPδ and C/EBPβ aligned with the consensus for SIAH2 interaction domains and two point mutations in C/EBPδ (underlined amino acids match the consensus motif). Immunoprecipitates with anti-Flag antibody and input were analyzed with antibodies specific for C/EBPβ and C/EBPδ and with anti-Flag to detect SIAH2. Cells had been treated with MG132 to stabilize C/EBPδ. (C) Western analysis of MCF-10A cells after transfection with the indicated C/EBPδ expression constructs and Flag-SIAH2. Fig 5. PMID:？22037769

(D) Western blot analysis with an anti-p24CA antibody detected the wild-type Gag protein pattern from the codon-optimized Hgpsyn？and truncated Gag isoforms (white arrows) encoded by UTRg*pRRE. Shown are results of two independent cotransfections with pcRev (lanes 2 and 3) and Hgpsyn？(lanes 4 and 5). Loading of similar amounts of protein was verified by incubation of the stripped Western blot with an antibody against the endogenous protein α-tubulin. Transfection efficiencies were similar, as demonstrated by a separate Western blot analysis against GFP encoded by the cotransfected pEGFP-C1. Fig 3. PMID:？22258250

Detection of nucleus-localized Gag. (A) HeLa cells transfected with the indicated？gag/gag-pol？expression plasmids were analyzed by confocal immunofluorescence microscopy. To demonstrate nuclear Gag signals, the contrast of anti-Gag immunofluorescence images was increased postimaging (Adobe Photoshop). White arrowheads mark examples of nuclei of untransfected cells that do not show nuclear Gag fluorescence after a contrast increase. White scale bars represent 10 μm. (B) Transfected HEK 293T cells were fractionated into nuclear (N) and cytoplasmic (C) fractions for anti-p24CA Western blot analysis. The stripped blot was incubated with an antibody against the cytoplasmic protein α-tubulin. Furthermore, the purity of the nuclear fractions was demonstrated by the lack of different cytoplasmic marker proteins. Fig 6. PMID:？22258250