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32518
TBR1 (D6C6X) Rabbit mAb (Alexa Fluor® 488 Conjugate)

TBR1 (D6C6X) Rabbit mAb (Alexa Fluor® 488 Conjugate) #32518

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
M Endogenous Rabbit IgG
IF-F

Confocal immunofluorescent analysis of postnatal day 5 (P5) mouse cortex using TBR1 (D6C6X) Rabbit mAb (Alexa Fluor® 488) (green) and Neurofilament-L (C28E10) Rabbit mAb (Alexa Fluor® 594 Conjugate) #8743 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Immunofluorescence (Frozen)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (9808) To prepare 1L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix. Adjust pH to 8.0.
  2. Formaldehyde: 16%, methanol free, Polysciences, Inc. (cat# 18814), use fresh and store opened vials at 4°C in dark, dilute in 1X PBS for use.
  3. Blocking Buffer (1X PBS / 5% normal goat serum (#5425) / 0.3% Triton™ X-100): To prepare 10 ml: add 0.5 ml normal goat serum and 0.5 ml 20X PBS to 9.0 ml dH2O, mix. While stirring, add 30 µl Triton™ X-100.
  4. Antibody Dilution Buffer: (1X PBS / 1% BSA / 0.3% Triton™ X-100): To prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml 1X PBS. Mix well then add 0.1 g BSA (#9998), mix.
  5. Prolong® Gold AntiFade Reagent (#9071), Prolong® Gold AntiFade Reagent with DAPI (#8961).

B. Specimen Preparation - Frozen/Cryostat Sections (IF-F)

  1. For fixed frozen tissue proceed with Immunostaining (Section C).
  2. For fresh, unfixed frozen tissue, please fix immediately, as follows:
    1. Cover sections with 4% formaldehyde dilute in 1X PBS.

      NOTE: Formaldehyde is toxic, use only in fume hood.

    2. Allow sections to fix for 15 minutes at room temperature.
    3. Aspirate liquid, rinse three times in 1X PBS for 5 minutes each.
    4. Proceed with Immunostaining (Section C).

C. Immunostaining

NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.

  1. Block specimen in Blocking Buffer for 60 min.
  2. While blocking, prepare primary antibody by diluting as indicated on datasheet in Antibody Dilution Buffer.
  3. Aspirate blocking solution, apply diluted primary antibody.
  4. Incubate overnight at 4°C.
  5. Rinse three times in 1X PBS for 5 min each.
  6. Coverslip slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961).
  7. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 4°C protected from light.

posted November 2006

revised November 2013

Protocol Id: 222

Application Dilutions
Immunofluorescence (Frozen) 1:50
Storage:

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

TBR1 (D6C6X) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total TBR1 protein. Low levels of nuclear staining of unknown specificity have been observed in mouse small intestine, spleen, and pancreas.

Species Reactivity:

Mouse

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro415 of human TBR1 protein.

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct immuno fluorescence analysis in mouse tissue. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated TBR1 (D6C6X) Rabbit mAb #49661.

T-box, brain, 1 (TBR1) is a transcription factor important in vertebrate embryo development. As a member of T-Box family of transcription factors, TBR1 is expressed in postmitotic glutamatergic projection neurons (1). During cortical neurogenesis, sequential expression of transcription factors Pax6, TBR2, and TBR1 regulates discrete steps in projection neuron differentiation (2). TBR1 is enriched in layer 6 of the developing cortex. In the absence of TBR1, TBR1 mutants exhibit profound defects in frontal cortex and layer 6 differentiation, suggesting that TBR1 regulates regional and laminar identity of postmitotic cortical neurons (3). Therefore, TBR1 expression can be used as a marker for postmitotic glutamatergic neurons and cortical laminar specificity.

  1. Hevner, R.F. et al. (2001) Neuron 29, 353-66.
  2. Englund, C. et al. (2005) J Neurosci 25, 247-51.
  3. Bedogni, F. et al. (2010) Proc Natl Acad Sci U S A 107, 13129-34.
Entrez-Gene Id
10716
Swiss-Prot Acc.
Q16650
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchased product solely in research, including use with HCS or other automated imaging applications but excluding use in combination with DNA microarrays. The buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

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To Purchase # 32518S

Product Number Size Price
32518S 100 µl (50 tests) $299.00.0
Quantity Subtotal
$0.00

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