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Purified Mouse Anti-MAP2B
Purified Mouse Anti-MAP2B

Western blot analysis of MAP2B on rat brain lysate. Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1:10000 dilution of MAP2B.

Purified Mouse Anti-MAP2B

Immunofluorescent staining of undifferentiated (left) and differentiated (right) SH-SY5Y cells.  Cells were seeded in a collagen coated 384 well imaging plate (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the anti-MAP2B antibody.  Differentiated cells were seeded in a 96 well, collagen coated imaging plate (Material # 353219) at ~ 5,000 cells per well.  Cells were incubated with 50 mM ATRA (Sigma, R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma, B3795) for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol, and the anti-MAP2B antibody.  The second step reagent in both cases was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen).  The images were taken on a Pathway 855 or 435 imager using a 20x objective.  This antibody also stained undifferentiated SK-N-SH cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Western blot analysis of MAP2B on rat brain lysate. Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1:10000 dilution of MAP2B.

Immunofluorescent staining of undifferentiated (left) and differentiated (right) SH-SY5Y cells.  Cells were seeded in a collagen coated 384 well imaging plate (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the anti-MAP2B antibody.  Differentiated cells were seeded in a 96 well, collagen coated imaging plate (Material # 353219) at ~ 5,000 cells per well.  Cells were incubated with 50 mM ATRA (Sigma, R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma, B3795) for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol, and the anti-MAP2B antibody.  The second step reagent in both cases was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen).  The images were taken on a Pathway 855 or 435 imager using a 20x objective.  This antibody also stained undifferentiated SK-N-SH cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure).

製品詳細
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BD Transduction Laboratories™
Rat (QC Testing), Human, Mouse (Tested in Development)
Mouse IgG1, κ
Human MAP2B aa. 19-219 Recombinant Protein
Western blot (Routinely Tested), Immunofluorescence, Immunohistochemistry (Tested During Development), Immunoprecipitation (Not Recommended)
280 kDa
250 µg/ml
AB_397833
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


推奨アッセイ手順

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

製品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
610460 Rev. 1
抗体の詳細
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18/MAP2B

Microtubule-associated proteins (MAPs) play a crucial role in the development and morphology of neurons. MAP2, specifically localized in dendrites, has four known isoforms that are produced by alternative splicing of the transcript and are expressed at various stages of neuronal development. MAP2B is a 280-kDa protein that is expressed throughout brain development. It contains functional domains that interact with the regulatory subunit of the cAMP-dependent kinase II and microtubules.  Experimental monitoring of its presence, along with GFAP and nestin, may be utilized to quantify neuronal development.

610460 Rev. 1
フォーマットの詳細
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
610460 Rev.1
引用&参考文献
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開発者向け参考資料 (2)

  1. Kanaani J, el-Husseini Ael-D, Aguilera-Moreno A, Diacovo JM, Bredt DS, Baekkeskov S. A combination of three distinct trafficking signals mediates axonal targeting and presynaptic clustering of GAD65. J Cell Biol. 2002; 158(7):1229-1238. (Clone-specific: Immunofluorescence). 参考文献を見る
  2. Kindler S, Schulz B, Goedert M, Garner CC. Molecular structure of microtubule-associated protein 2b and 2c from rat brain. J Biol Chem. 1990; 265(32):19679-19684. (Biology). 参考文献を見る
610460 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.