Anti-alpha Tubulin antibody - Loading Control (ab4074)
Key features and details
- Rabbit polyclonal to alpha Tubulin - Loading Control
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Rat, Chicken, Cow, Human, Chinese hamster
- Isotype: IgG
Overview
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Product name
Anti-alpha Tubulin antibody - Loading Control
See all alpha Tubulin primary antibodies -
Description
Rabbit polyclonal to alpha Tubulin - Loading Control -
Host species
Rabbit -
Specificity
This antibody detects a single band in cell lines, corresponding to alpha tubulin. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB MouseHuman
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Immunogen
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Positive control
- WB: HeLa, NIH 3T3, and PC12 cell lysates. ICC/IF: HeLa cells. IHC-P: Human endometrium tissue.
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General notes
This antibody makes an excellent loading control.
Images
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Western blot - Anti-alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
Lane 2 : NIH/3T3 (Mouse) Whole Cell Lysate (ab52956)
Lane 3 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab4074) Image from PLoS One. 2014; 9(10): e109590. Fig 5o,doi: 10.1371/journal.pone.0109590 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Immunocytochemistry of SisNPFR-C6E8 cell line. Cells were prefixed in freshly prepared 4% paraformaldehyde (PFA) in F12 K culture medium with 10% fetal bovine serum and Geneticin. Fixed cells were permeabilized in PBST (0.25% Triton X-100 in PBS) for 5 min and blocked at 4°C overnight in 5% normal goat serum. The cells were then incubated at 4°C overnight with ab4074 (1:50). Cover slips were placed onto slides and mounted with VectaShield containing DAPI.
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Western blot - Anti-alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
Lane 2 : HEK293 (Human) Whole Cell Lysate (ab52256)
Lane 3 :NIH/3T3 whole cell lysate (ab7179)
Lane 4 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lane 6 : HEK293 (Human) Whole Cell Lysate (ab52256) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lane 7 :NIH/3T3 whole cell lysate (ab7179) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lane 8 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab4074)ICC/IF image of ab4074 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4074, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red). -
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : Mouse 3T3 lysate
Lane 2 : Rat liver lysate
Lane 3 : Cow cell lysate
Lane 4 : Chicken cell lysate
Lane 5 : CHO lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 30 secondsab4074 cross-reacts strongly with Mouse 3T3 lysate and shows weak cross-reactivity with Rat liver, Cow, Chicken and CHO lysates.
