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beta-III Tubulin Antibody - BSA Free

Novus Biologicals | Catalog # NB100-1612

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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry Whole-Mount, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry Whole-Mount, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Chicken IgY

Format

BSA Free
View all beta-III Tubulin Primary Antibodies »

Product Specifications

Immunogen

Chickens were immunized with three synthetic peptide/keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of beta-III Tubulin, but are shared between the human (NP_AAL28094, NCBI) and rat (AAM28438, NCBI) protein sequences.

Marker

Neuron Cell Marker

Clonality

Polyclonal

Host

Chicken

Isotype

IgY

Theoretical MW

50 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for beta-III Tubulin Antibody - BSA Free

Western Blot: beta-III Tubulin Antibody [NB100-1612]

Western Blot: beta-III Tubulin Antibody [NB100-1612]

beta-III-Tubulin-Antibody-Western-Blot-NB100-1612-img0006.jpg
Immunocytochemistry/ Immunofluorescence: beta-III Tubulin Antibody [NB100-1612]

Immunocytochemistry/ Immunofluorescence: beta-III Tubulin Antibody [NB100-1612]

Immunocytochemistry/Immunofluorescence: beta-III Tubulin Antibody [NB100-1612] - Beta-III Tubulin in rat primary motor neurons. Ab dilution 1:1000 in PBST (0.1% Triton X-100) + 10% GS O/N at 4C. ICC/IF image submitted by a verified customer review.
Western Blot: beta-III Tubulin Antibody [NB100-1612]

Western Blot: beta-III Tubulin Antibody [NB100-1612]

beta-III-Tubulin-Antibody-Western-Blot-NB100-1612-img0007.jpg
beta-III Tubulin Antibody Western Blot: beta-III Tubulin Antibody [NB100-1612]

Western Blot: beta-III Tubulin Antibody [NB100-1612]

beta-III Tubulin Western Blot using homogenates of adult mouse brain (1:3000).
beta-III Tubulin Antibody

Western Blot: beta-III Tubulin Antibody [NB100-1612] -

Western Blot: beta-III Tubulin Antibody [NB100-1612] - The levels of glial fibrillary acidic protein (GFAP) & S100 beta as astrocytic protein marker proteins in the hippocampal area. (A) Western blot images using GFAP, tubulin beta‐3, & beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized GFAP/ beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White & black bars indicate the wild‐type C57BL/6J & Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P < 0.05; **P < 0.01; n = >12 in each group. (B) Western blot images using S100 beta, tubulin beta‐3, & beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized S100 beta / beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White & black bars indicate the wild‐type C57BL/6J & Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P < 0.05; **P < 0.01; n = >12 in each group. (C) Micrographs of immunohistochemical analysis on paraffined hippocampal tissue sections using GFAP & S100 beta antibody. Brown cells indicate GFAP‐stained astrocytes. Scale bar = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27623715), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for beta-III Tubulin Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:2000-1:5000

Immunohistochemistry

1:2000 - 1:5000

Western Blot

1:10000 - 1:20000
Application Notes
Although not tested, this antibody may be useful for immunohistochemistry on frozen sections. Use in KD reported in scientific literature (PMID:33333046). Each of the three antibodies were analyzed by immunohistochemistry (1:2000). using fluorescein-labeled goat anti-chicken IgY (1:500). as the secondary reagent. Use in Immunohistochemistry-Whole mount reported in scientific literature (PMID 27178445). Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID: 27178445). Use in Flow Cytometry reported in scientific literature (PMID: 31755958).

Reviewed Applications

Read 2 reviews rated 4.5 using NB100-1612 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

10mM PBS (0.9% isotonic, w/v, pH 7.2)

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

0.3 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: beta-III Tubulin

Beta-III Tubulin is abundant in the central and peripheral nervous systems where it is prominently expressed during fetal and postnatal development. Beta III Tubulin is often referred to as TUJ1 because Tuj-1 is the clone name of the original monoclonal antibody. The Tuj1 clone is such a popular neuronal marker that Tuj-1 is now used as a synonym for beta III tubulin. The distribution of Tubulin Beta 3 is neuron-associated, exhibiting distinct temporospatial gradients according to the regional neuroepithelia of origin. However, transient expression of the Tubulin Beta 3 protein is also present in the subventricular zones of the CNS comprising putative neuronal- and/or glial precursor cells, as well as in Kulchitsky neuroendocrine cells of the fetal respiratory epithelium. This temporally restricted, potentially non-neuronal expression may have implications in the identification of presumptive neurons derived from embryonic stem cells. Mutation on beta-tubulin has been linked to anti-cancer drug resistance in lung cancers.

Alternate Names

beta-Tubulin III, Beta3-tubulin, betaIII Tubulin, Class III beta-tubulin, TUBB3, Tubulin Beta 3, Tubulin beta-4, Tubulin beta-III, TUJ1 antigen, TUJ-1 antigen

Entrez Gene IDs

10381 (Human); 246118 (Rat)

Gene Symbol

TUBB3

UniProt

Q13509 (Human)

Additional beta-III Tubulin Products

Product Documents for beta-III Tubulin Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for beta-III Tubulin Antibody - BSA Free

Chicken products cannot be exported to Canada.

Purification Notes
After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 300 ug/ml. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

Storage Notes
Store at 4C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for beta-III Tubulin Antibody - BSA Free

Customer Reviews for beta-III Tubulin Antibody - BSA Free (2)

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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • beta-III Tubulin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Rat Primary Motor Neurons
    Species: Rat
    Verified Customer | Posted 02/19/2020
    beta-III Tubulin in rat primary motor neurons. Ab dilution 1:1000 in PBST (0.1% triton) + 10% GS O/N 4ºC.
    Fixed in PFA 4% + Sacharose 4% in PBS 20 min RT. Ab dilution 1:1000 in PBST (0.1% triton) + 10% GS O/N 4ºC.
    beta-III Tubulin Antibody - BSA Free NB100-1612
  • Name: Paula Loos
    Application: Immunocytochemistry
    Sample Tested:
    Species: Rat
    Verified Customer | Posted 05/19/2014
    E18 rat primary hippocampal neurons 8 DIV. beta tubulin III
    beta-III Tubulin Antibody - BSA Free NB100-1612

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Protocols

View specific protocols for beta-III Tubulin Antibody - BSA Free (NB100-1612):


Citrate Buffer Antigen Retrieval Protocol

Background: Formaldehyde fixation (2% or 4%, or as a component of 10% formalin) produces protein cross-links in tissues that tends to interfere with antibody penetration. This seems to be particularly true of paraffin- embedded formaldehyde-fixed tissue. Since chicken IgY antibodies are larger than rabbit or mouse IgG's, "extra steps" may be necessary to compensate for their larger size.

The citrate-based "antigen retrieval" protocol outlined below has been shown to improve chicken IgY antibody penetration into 4% formalde- hyde-fixed paraffin-embedded sections, and can increase the degree and intensity of immunoreactivity and immunostaining.

Reagents (NOTE: You can use either the Sodium Citrate or Citric Acid Buffers in step #3, below)

"Sodium Citrate Buffer" (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0)

Weigh out 2.94 grams of trisodium citrate (dihydrate). Dissolve in approximately 900 mls of deionized, distilled water. Adjust the pH to 6.00 with 1.0 N HCl. Add
0.5 ml of Tween-20. Mix. Bring up the volume to 1.0 litres with water. Store this solution at room temperature for 3 months or at 4C for longer periods.

"Citric Acid Buffer" (10mM Citric Acid, 0.05% Tween 20, pH 6.0)

Weigh out 1.92 grams of citric acid (anhydrous). Dissolve in approximately 900 mls of deionized, distilled water. Adjust the pH to 6.0 with 1.0 N NaOH. Add
0.5 ml of Tween-20. Mix. Bring up the volume to 1.0 litres with water. Store this solution at room temperature for 3 months or at 4C for longer periods.

"Phosphate-Buffered Saline" [PBS, 10 mM Sodium phosphate-buffered (pH 7.2) isotonic (0.9%, w/v) saline solution] PBS Tween (0.05% Tween 20 in PBS)
Ethanol (80%, 90%, 95%, 100%) diluted with water.

Xylene

Procedure (for use with paraffin-embedded sections):

1 Deparaffinize tissue sections in 2 changes of xylene (5 minutes each).

2. Hydrate in 2 changes of 100% ethanol (3 minutes each), 95% ethanol (1 minute), 90% ethanol (1 minute), 80% ethanol (1 minute). Rinse in distilled water.

3. Pre-heat steamer or water bath with staining dish containing either Sodium Citrate Buffer or Citrate Buffer. Wait until temperature reaches 95-100 degrees C.

NOTE: Microwave or pressure cooker can be used as an alternative as a heating source.

4. Immerse slides in the staining dish. Place the lid loosely on the staining dish and incubate for 20-40 minutes (optimal incubation times will vary).

5. Remove the staining dish, and allow it to cool to room temperature (for 20 minutes or so).

6. Rinse sections in PBS Tween twice for 2 minutes each time.

NOTE: The remainder of this protocol is meant to be a suggestion, and can be substituted with your regular immunostaining protocol.

7. Block sections for 30 minutes with Blocking buffer diluted 1:10 with water.

8. Incubate sections with primary antibody at appropriate dilution in antibody dilution buffer overnight at 4 degrees C. Since chicken IgY antibodies are larger than mammalian IgG's, this overnight incubation allows more time for antibody penetration into tissue sections.

9. Rinse sections with PBS Tween 20 twice for 5 minutes each time.

10. Incubate sections with labeled secondary antibody (see NOTE, below) at appropriate dilution (for one hour at room temperature) in a 1:100 dilution of blocking buffer (diluted in PBS).

11. Rinse with PBS Tween 20 for three times for 5 minutes each time.

NOTE: This protocol may use HRP- or fluorescently-labeled secondary antibodies produced in goats or rabbits.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for beta-III Tubulin Antibody - BSA Free

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  • Q: Do you know if the beta-III tubulin antibody (Cat NB100-1612) has been successfully used for ELISA?

    A: Our tubulin beta 3 antibody with catalogue number NB100-1612 has not yet been validated for ELISA.

  • Q: I would like to find out the isotype information for Catalog Number NB100-1612. Is it IgG or IgY? What kind of secondary antibody should I use?

    A: Our tubulin beta 3 antibody with catalogue number NB100-1612 is a chicken polyclonal, of the isotype IgY. You can see our relevant secondary antibodies for the detection of chicken IgY at the following link: View Secondary Antibodies

  • Q: Do you know if the beta-III tubulin antibody (Cat NB100-1612) has been successfully used for ELISA?

    A: Our tubulin beta 3 antibody with catalogue number NB100-1612 has not yet been validated for ELISA.

  • Q: I would like to find out the isotype information for Catalog Number NB100-1612. Is it IgG or IgY? What kind of secondary antibody should I use?

    A: Our tubulin beta 3 antibody with catalogue number NB100-1612 is a chicken polyclonal, of the isotype IgY. You can see our relevant secondary antibodies for the detection of chicken IgY at the following link: View Secondary Antibodies

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