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Ab72439

Manufactured by Abcam
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Ab72439 is a lab equipment product offered by Abcam. It is designed for use in scientific research applications. The core function of this product is to [CORE FUNCTION]. No further details or interpretations are provided.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) M mlv rt system, by Promega (1 mentions) Rabbit anti β actin, by Cell Signaling Technology (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Mouse anti flag, by Merck Group (1 mentions) Mab318, by Merck Group (1 mentions) Anti gfp antibody, by Abcam (1 mentions) Anti glial fibrillary acidic protein antibody, by Merck Group (1 mentions) Human bdnf elisa kit, by Abbexa (1 mentions) Anti neun antibody, by Abcam (1 mentions) Ab6160, by Abcam (1 mentions) Ab106814, by Abcam (1 mentions) Ab12274, by Abcam (1 mentions) Ab32127, by Abcam (1 mentions) Mitoprofile total oxphos rodent wb antibody cocktail, by Abcam (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions)

4 protocols using ab72439

1

Quantifying mRNA and Protein Expression

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Total RNA was extracted from tissues or cultured cells using TRIzol® (Invitrogen), and cDNA was synthesized using M-MLV RT System (Promega) followed by PCR amplification and quantification using SYBR® Green PCR Master Mix (Applied Biosystems). House-keeping β-actin mRNA was used for PCR normalization. Animal tissues were homogenized, and proteins were dissolved in a lysis buffer. Proteins dissolved in lysis buffer were separated by SDS-PAGE and identified by immunoblotting. Primary antibodies included rabbit anti-p-RelA (3033, Cell Signaling), rabbit anti-RelA (8242, Cell Signaling), rabbit anti-β-actin (4970, Cell Signaling), mouse anti-flag (F1804, Sigma), rabbit anti-BDNF (ab72439, Abcam) and rabbit anti-GAT3 (ab431, Abcam). Secondary antibodies included HRP-conjugated anti-mouse and anti-rabbit antibodies (Pierce).
Zhang Y., Reichel J.M., Han C., Zuniga-Hertz J.P, & Cai D. (2017). Astrocytic process plasticity and IKKβ/NF-κB in central control of blood glucose, blood pressure and body weight. Cell metabolism, 25(5), 1091-1102.e4.
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2

Immunohistochemical Analysis of Ovarian Cancer

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Both frozen and paraffin-embedded sections were used for immunohistochemistry. Paraffin-embedded sections of human ovarian cancer were deparaffinized in xylene and dehydrated-rehydrated in alcohol and PBS. Antigen retrieval was performed using citrate buffer in the microwave for ten minutes. Frozen sections were fixed in acetone and acetone-chloroform. After endogenous peroxide blocking and three washes with PBS, the slides were incubated with tyrosine hydroxylase antibody (MAB318, 1:50; Millipore), BDNF (ab72439, 1:100, Abcam) or Neurofilament antibody overnight at 4°C. Matching secondary antibodies were used for one hour at room temperature and staining was developed using DAB. Hematoxylin was used to stain nuclei. Nerves within tumors were quantified based on presence of both individual nerves and bundles that stained positive for NF or TH.
Allen J.K., Armaiz-Pena G.N., Nagaraja A.S., Sadaoui N.C., Ortiz T., Dood R., Ozcan M., Herder D.M., Haemerrle M., Gharpure K.M., Rupaimoole R., Previs R.A., Wu S.Y., Pradeep S., Xu X., Han H.D., Zand B., Dalton H.J., Taylor M., Hu W., Bottsford-Miller J., Moreno-Smith M., Kang Y., Mangala L.S., Rodriguez-Aguayo C., Sehgal V., Spaeth E.L., Ram P.T., Wong S.T., Marini F.C., Lopez-Berestein G., Cole S.W., Lutgendorf S.K., De Biasi M, & Sood A.K. (2018). Sustained adrenergic signaling promotes intratumoral innervation through BDNF induction. Cancer research, 78(12), 3233-3242.
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3

Evaluating Human BDNF Expression in P301L Mouse Brains

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ELISA, western blot and immunohistochemistry assays were performed to examine the expression of human BDNF in the brains of P301L mice after intraventricular injection of AAV-BDNF. Fresh left hemisphere was weighed and homogenized in lysate buffer. The homogenates were centrifuged at 10  000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa). In addition, the levels of total BDNF including endogenous mouse BDNF and human BDNF in brain homogenates were also detected by western blot analysis using anti-BDNF antibody (Abcam, ab72439, detecting both mouse and human BDNF). Coronal sections from the right hemisphere were immunostained for microglia (anti-CD45 antibody, Millipore, Bedford, MA, USA), astrocyte (anti-glial fibrillary acidic protein antibody, Millipore), neurons (anti-NeuN antibody, Abcam), GFP (anti-GFP antibody, Abcam) and human BDNF (anti-human BDNF antibody, R&D systems, Minneapolis, MN, USA) to identify localization and levels of human BDNF expression in P301L mouse brains after AAV-BDNF injection.
Jiao S.S., Shen L.L., Zhu C., Bu X.L., Liu Y.H., Liu C.H., Yao X.Q., Zhang L.L., Zhou H.D., Walker D.G., Tan J., Götz J., Zhou X.F, & Wang Y.J. (2016). Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease. Translational Psychiatry, 6(10), e907-.
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4

Western Blot Analysis of Brain Proteins

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Western blot analysis was carried out in brain tissue homogenates as reported previously [36 (link)]. Briefly, brain samples were homogenized in lysis buffer. Total protein content was determined using a Pierce™ BCA Protein Assay Kit (Thermo Scientific). Band analysis was performed using ChemiDoc XRS system (BioRad, Munich, Germany). The following antibodies were used: full-length AβPP (fl-AβPP) and C-terminal fragments (CTFs) were detected using AβPP C-Terminal Fragment (C1/6.1) antibody (SIG-39152; Covance, Princeton, NJ, USA). Detection of respiratory system complexes was carried out by MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail (ab110413; abcam, Cambridge, UK). As primary antibody for synaptic markers, Anti-BDNF (ab72439; abcam), Anti-GAP43 (ab12274; abcam), and Anti-synaptophysin (ab32127; abcam) were used. To detect protein expression of PGC-1α in the brain, Anti-PGC-1α antibody (ab106814; abcam) was used. Anti-tubulin antibody (ab6160; abcam) and Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (MAB374; Merck Millipore, Darmstadt, Germany) were used to verify equal protein loading.
Pohland M., Pellowska M., Asseburg H., Hagl S., Reutzel M., Joppe A., Berressem D., Eckert S.H., Wurglics M., Schubert‐Zsilavecz M, & Eckert G.P. (2018). MH84 improves mitochondrial dysfunction in a mouse model of early Alzheimer’s disease. Alzheimer's Research & Therapy, 10, 18.
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