Anti-alpha Tubulin 抗体 - Loading Control (ab4074)

製品の概要

  • 製品名Anti-alpha Tubulin antibody - Loading Control
    alpha Tubulin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to alpha Tubulin - Loading Control
  • 特異性This antibody detects a single band in cell lines, corresponding to alpha tubulin.
  • アプリケーション適用あり: IHC-P, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Cow, Human
    交差が予測される動物種: Pig, Xenopus laevis, Chimpanzee, Monkey, Zebrafish, Chinese Hamster
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human alpha Tubulin.

    (Peptide available as ab23537.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in HeLa cells in WB and IF.
  • 特記事項


    This antibody makes an excellent loading control.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4074 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). Can be blocked with alpha Tubulin peptide (ab23537).
ICC/IF Use a concentration of 1 µg/ml.

ターゲット情報

  • 関連性Microtubules are involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintenance of cell shape. Tubulin itself is a globular protein which consists of two polypeptides (alpha and beta tubulin). Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter. Tyrosine ligase adds a C-terminal tyrosin to monomeric alpha tubulin. Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated alpha tubulin is referred to as Glu-tubulin. Another post-translational modification of detyrosinated alpha tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle. Alpha tubulin is not suitable as a loading control in adipose tissue as expression of tubulin in adipose tissue is very low ( Spiegelman and Farmer, Cell, 1982, 29(1): 53-60, "in cells undergoing adipose differentiation actin synthesis decreases by 90%").
  • 細胞内局在Major constituent of Microtubules
  • 参照データベース
  • 別名
    • TUBA1 antibody
    • TUBA1A antibody
    • TUBA2 antibody
    • TUBA3 antibody
    • TUBA3C antibody
    • TUBA4A antibody
    • Tubulin, alpha 1a antibody
    • Tubulin, alpha 3c antibody
    • Tubulin, alpha 4a antibody
    see all

Anti-alpha Tubulin antibody - Loading Control 画像

  • 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 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)

    Lysates/proteins at 10 µg per lane.

    Secondary
    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 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.

  • 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 : PC12 (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 : NIH 3T3 whole cell lysate (ab7179) 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 : PC12 (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
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 50 kDa
  • (Image courtesy of Human Protein Atlas).
    ab4074 staining alpha tubulin in the glandular cells of the endometrium. Paraffin embedded endometrium tissue of a 33 year-old female was incubated with ab4074 (1/25 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab4074 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .



  • Predicted band size : 50 kDa
  • 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). 
  • 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
    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 seconds

    ab4074 cross-reacts strongly with Mouse 3T3 lysate and shows weak cross-reactivity with Rat liver, Cow, Chicken and CHO lysates.

Anti-alpha Tubulin antibody - Loading Control (ab4074) 使用論文

This product has been referenced in:
  • Elias I  et al. ALOX5AP Overexpression in Adipose Tissue Leads to LXA4 Production and Protection Against Diet-Induced Obesity and Insulin Resistance. Diabetes 65:2139-50 (2016). Read more (PubMed: 27207555) »
  • El-Sikhry HE  et al. Novel Roles of Epoxyeicosanoids in Regulating Cardiac Mitochondria. PLoS One 11:e0160380 (2016). WB ; Human . Read more (PubMed: 27494529) »

See all 70 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (Ovarian Carcinoma (TOV112D))
Specification Ovarian Carcinoma (TOV112D)
Permeabilization Yes - 0.1% Triton-x100
Fixative Paraformaldehyde
Username

Dr. Gajendra Katara

Verified customer

投稿 Aug 18 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (SKOV-3 Ovarian Carcinoma)
Specification SKOV-3 Ovarian Carcinoma
Permeabilization Yes - 0.1% Triton-x100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Mouse Cell lysate - whole cell (Mouse embryonic stem cells)
Specification Mouse embryonic stem cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Jul 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (mouse fibroblast)
Loading amount 50 µg
Specification mouse fibroblast
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Mar 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (ovarian cancer cells)
Permeabilization Yes - methanol:acetone (9:1)
Specification ovarian cancer cells
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Dr. Gajendra Katara

Verified customer

投稿 Feb 15 2016

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Hep G2, liver)
Specification Hep G2, liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Mar 17 2015

Thank you for your answer and clarification. This is an extremely important piece of information.

The most widely used loading controls are tubulin, actin and GAPDH. However, some chemicals can alter the expression levels of these proteins so...

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Thank your for your enquiry and interest in our products.

Could you please explain what experimental system you use with the mouse liver samples? Do you apply any drug which may alter protein expression?

I look forward to hearing fr...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human fibroblast cell)
Loading amount 50 µg
Specification human fibroblast cell
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Nov 28 2012

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this h...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"