Overview

  • Product name
    Anti-Tubulin antibody [DM1A +DM1B] - Loading Control
    See all Tubulin primary antibodies
  • Description
    Mouse monoclonal [DM1A +DM1B] to Tubulin - Loading Control
  • Host species
    Mouse
  • Tested applications
    Suitable for: Electron Microscopy, ICC/IF, IP, IHC-P, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Drosophila melanogaster
    Predicted to work with: Chicken, Guinea pig, Cow, Pig, Gerbil
  • Immunogen

    Tissue/ cell preparation, Native chick brain microtubules.

  • Epitope
    aa 426-450
  • Positive control
    • In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; NIH3T3; PC12. Ls174T, MAD109 cells. Skin or lung.
  • General notes

    Please note that this antibody is an oligoclonal antibody. It is a cocktail of monoclonal antibodies that have been carefully selected. Oligoclonal antibodies have not only the specificity and batch-to-batch consistency of a monoclonal antibody, but also have the advantage of the sensitivity of a polyclonal antibody due to their ability to recognize multiple epitopes on an antigen. 

Properties

Applications

Our Abpromise guarantee covers the use of ab44928 in the following tested applications.

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

Application Abreviews Notes
Electron Microscopy Use at an assay dependent concentration.
ICC/IF 1/2000.
IP 1/400.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 50 kDa).
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha-chain.
  • Sequence similarities
    Belongs to the tubulin family.
  • Post-translational
    modifications
    Undergoes a tyrosination/detyrosination cycle, the cyclic removal and re-addition of a C-terminal tyrosine residue by the enzymes tubulin tyrosine carboxypeptidase (TTCP) and tubulin tyrosine ligase (TTL), respectively.
    Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
    Acetylation of alpha-tubulins at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
  • Cellular localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Tubulin beta 2b antibody
    • Alpha tubulin antibody
    • Alpha-tubulin ubiquitous antibody
    • beta Ib tubulin antibody
    • CDCBM5 antibody
    • CDCBM6 antibody
    • fd02b12 antibody
    • K ALPHA 1 antibody
    • M40 antibody
    • OK/SW-cl.56 antibody
    • TBA1B_HUMAN antibody
    • TUBA1B antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin alpha 1b antibody
    • Tubulin alpha-1B chain antibody
    • Tubulin alpha-ubiquitous chain antibody
    • Tubulin beta 1b antibody
    • Tubulin beta 2A antibody
    • tubulin beta 2A class IIa antibody
    • Tubulin beta antibody
    • tubulin beta chain antibody
    • tubulin beta class I antibody
    • tubulin beta-1 chain antibody
    • tubulin beta-2A chain antibody
    • tubulin beta-5 chain antibody
    • Tubulin K-alpha-1 antibody
    • tubulin, alpha, ubiquitous antibody
    • tubulin, beta 2A class IIa antibody
    • tubulin, beta polypeptide 2 antibody
    • tubulin, beta polypeptide antibody
    • tubulin, beta, class IIA antibody
    • wu:fd02b12 antibody
    • zgc:55461 antibody
    see all

Images

  • Ab44928 staining human normal placenta. Staining is localized to the cytoplamsm
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ab44928 staining Tubulin in Drosophila S2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol and blocked with 5% BSA for 15 minutes at Room temperature. Samples were incubated with primary antibody (1/2000 in 5% BSA + PBST) for 2 hours. Ab6785 (1/1000) was used as secondary antibody.

    See Abreview

  • All lanes : Anti-Tubulin antibody [DM1A +DM1B] - Loading Control (ab44928) at 1/5000 dilution

    All lanes : Whole cell lysate prepared from human colon cells

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP sheep anti-mouse polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 50 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute

    See Abreview

  • All lanes : Anti-Tubulin antibody [DM1A +DM1B] - Loading Control (ab44928) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 53 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes
  • Overlay histogram showing HeLa cells stained with ab44928 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab449287, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Popp I  et al. Fanconi anemia with sun-sensitivity caused by a Xeroderma pigmentosum-associated missense mutation in XPF. BMC Med Genet 19:7 (2018). Read more (PubMed: 29325523) »
  • Ruiz de Garibay G  et al. Tumor xenograft modeling identifies an association between TCF4 loss and breast cancer chemoresistance. Dis Model Mech 11:N/A (2018). Read more (PubMed: 29666142) »
See all 20 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Zebrafish Cell lysate - whole cell (brain tissue)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
5 µg
Specification
brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Tyrone Genade

Verified customer

Submitted Jun 19 2015

Application
Western blot
Sample
Nothobranchius furzeri Cell lysate - whole cell (brain)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
5 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Tyrone Genade

Verified customer

Submitted Jun 18 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample
Human Tissue sections (Left atrial appendage)
Specification
Left atrial appendage
Permeabilization
No
Fixative
Paraformaldehyde

Dr. Beth Bunnell

Verified customer

Submitted Sep 08 2014

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-20%)
Sample
Human Cell lysate - other (A375 melanoma cells)
Specification
A375 melanoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jun 10 2013

Answer

Thank you for these details.

I appreciate the time you have spent on these experiments and would be pleased to arrange a credit note or a replacement with a different lot number of ab44928 or with an alternative anti-tubulin form our catalogue. The link to the list of Abcam anti-tubulin antibodies is: https://www.abcam.com/index.html?t=194&pt=1.

I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for contacting us.

I am not sure I understand the problem you are experiencing with the anti-tubulin ab44928.
Are you observing an extra band in Western Blot after immunoprecipitation?
What type of sample are you working on?
Do you have an image of your blot so I can have a better understanding of the problem?
Have you run a "no-primary" control to identify if the unspecific binding comes from the secondary or the primary?

I look forward to your reply.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products.
We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

In order to trace the lot number as well as the order numberrelated to the antibody ab44928, could you also please send me the date of purchase and inform me if you placed your order via our local distributor?

I look forward to receiving your reply.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (retina or eyecup without retina, also rat cells in)
Loading amount
20 µg
Specification
retina or eyecup without retina, also rat cells in
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Colon)
Loading amount
20 µg
Specification
Colon
Gel Running Conditions
Reduced Denaturing (12.5)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Mr. Qifeng Lin

Verified customer

Submitted Jul 11 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cell (Drosophila S2 cells)
Specification
Drosophila S2 cells
Fixative
Methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: R.T°C

Ms. Maureen Sinclair

Verified customer

Submitted Feb 18 2010

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

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