Structured Review

PerkinElmer ht22 cells
Schematic illustration of the role of JNK-p53-GADD45α signalling cascade in mediating the oxidative cytotoxicity in <t>HT22</t> neuronal cells. During glutamate-induced oxidative stress, JNK is activated first as a result of ROS accumulation, and the
Ht22 Cells, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 1503 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 1503 article reviews
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ht22 cells - by Bioz Stars, 2020-09
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1) Product Images from "Critical role of the JNK-p53-GADD45? apoptotic cascade in mediating oxidative cytotoxicity in hippocampal neurons"

Article Title: Critical role of the JNK-p53-GADD45? apoptotic cascade in mediating oxidative cytotoxicity in hippocampal neurons

Journal: British Journal of Pharmacology

doi: 10.1111/j.1476-5381.2010.01041.x

Schematic illustration of the role of JNK-p53-GADD45α signalling cascade in mediating the oxidative cytotoxicity in HT22 neuronal cells. During glutamate-induced oxidative stress, JNK is activated first as a result of ROS accumulation, and the
Figure Legend Snippet: Schematic illustration of the role of JNK-p53-GADD45α signalling cascade in mediating the oxidative cytotoxicity in HT22 neuronal cells. During glutamate-induced oxidative stress, JNK is activated first as a result of ROS accumulation, and the

Techniques Used:

GADD45α contributes to glutamate-induced oxidative cytotoxicity in HT22 cells. After the HT22 cells were incubated with 5 mmol·L −1 glutamate for 6, 12 and 24 h, they were subjected to analysis of the GADD45α mRNA levels
Figure Legend Snippet: GADD45α contributes to glutamate-induced oxidative cytotoxicity in HT22 cells. After the HT22 cells were incubated with 5 mmol·L −1 glutamate for 6, 12 and 24 h, they were subjected to analysis of the GADD45α mRNA levels

Techniques Used: Incubation

JNK activation contributes to glutamate-induced cell death (A, upper panels). HT22 cells were first incubated with 5 mmol·L −1 glutamate for 3, 6, 12 and 24 h, and then the cellular extracts were subjected to Western blotting for the levels
Figure Legend Snippet: JNK activation contributes to glutamate-induced cell death (A, upper panels). HT22 cells were first incubated with 5 mmol·L −1 glutamate for 3, 6, 12 and 24 h, and then the cellular extracts were subjected to Western blotting for the levels

Techniques Used: Activation Assay, Incubation, Western Blot

Knock-down of p53 or GADD45α suppresses glutamate-induced apoptosis. (A) HT22 cells were first transfected with the negative control siRNA (siCon), p53 siRNA (sip53), GADD45α siRNA (siGADD45α), or a mock transfection procedure
Figure Legend Snippet: Knock-down of p53 or GADD45α suppresses glutamate-induced apoptosis. (A) HT22 cells were first transfected with the negative control siRNA (siCon), p53 siRNA (sip53), GADD45α siRNA (siGADD45α), or a mock transfection procedure

Techniques Used: Transfection, Negative Control

Knock-down of p53 suppresses glutamate-induced GADD45α expression. (A, left panel) HT22 cells were transfected with either the negative control siRNA (siCon) or p53 siRNA (sip53), and 24 h later, cells were exposed to 5 mmol·L −1
Figure Legend Snippet: Knock-down of p53 suppresses glutamate-induced GADD45α expression. (A, left panel) HT22 cells were transfected with either the negative control siRNA (siCon) or p53 siRNA (sip53), and 24 h later, cells were exposed to 5 mmol·L −1

Techniques Used: Expressing, Transfection, Negative Control

Glutamate induces p53 activation in HT22 cells. (A) HT22 cells were incubated with 5 mmol·L −1 glutamate for the indicated length of time. Whole cell extracts were examined by Western blotting using the anti-phospho-p53 (Ser15) and anti-p53
Figure Legend Snippet: Glutamate induces p53 activation in HT22 cells. (A) HT22 cells were incubated with 5 mmol·L −1 glutamate for the indicated length of time. Whole cell extracts were examined by Western blotting using the anti-phospho-p53 (Ser15) and anti-p53

Techniques Used: Activation Assay, Incubation, Western Blot

2) Product Images from "Critical role of the JNK-p53-GADD45? apoptotic cascade in mediating oxidative cytotoxicity in hippocampal neurons"

Article Title: Critical role of the JNK-p53-GADD45? apoptotic cascade in mediating oxidative cytotoxicity in hippocampal neurons

Journal: British Journal of Pharmacology

doi: 10.1111/j.1476-5381.2010.01041.x

GADD45α contributes to glutamate-induced oxidative cytotoxicity in HT22 cells. After the HT22 cells were incubated with 5 mmol·L −1  glutamate for 6, 12 and 24 h, they were subjected to analysis of the GADD45α mRNA levels
Figure Legend Snippet: GADD45α contributes to glutamate-induced oxidative cytotoxicity in HT22 cells. After the HT22 cells were incubated with 5 mmol·L −1 glutamate for 6, 12 and 24 h, they were subjected to analysis of the GADD45α mRNA levels

Techniques Used: Incubation

JNK activation contributes to glutamate-induced cell death (A, upper panels). HT22 cells were first incubated with 5 mmol·L −1  glutamate for 3, 6, 12 and 24 h, and then the cellular extracts were subjected to Western blotting for the levels
Figure Legend Snippet: JNK activation contributes to glutamate-induced cell death (A, upper panels). HT22 cells were first incubated with 5 mmol·L −1 glutamate for 3, 6, 12 and 24 h, and then the cellular extracts were subjected to Western blotting for the levels

Techniques Used: Activation Assay, Incubation, Western Blot

Knock-down of p53 suppresses glutamate-induced GADD45α expression. (A, left panel) HT22 cells were transfected with either the negative control siRNA (siCon) or p53 siRNA (sip53), and 24 h later, cells were exposed to 5 mmol·L −1
Figure Legend Snippet: Knock-down of p53 suppresses glutamate-induced GADD45α expression. (A, left panel) HT22 cells were transfected with either the negative control siRNA (siCon) or p53 siRNA (sip53), and 24 h later, cells were exposed to 5 mmol·L −1

Techniques Used: Expressing, Transfection, Negative Control

Glutamate induces p53 activation in HT22 cells. (A) HT22 cells were incubated with 5 mmol·L −1  glutamate for the indicated length of time. Whole cell extracts were examined by Western blotting using the anti-phospho-p53 (Ser15) and anti-p53
Figure Legend Snippet: Glutamate induces p53 activation in HT22 cells. (A) HT22 cells were incubated with 5 mmol·L −1 glutamate for the indicated length of time. Whole cell extracts were examined by Western blotting using the anti-phospho-p53 (Ser15) and anti-p53

Techniques Used: Activation Assay, Incubation, Western Blot

Related Articles

Amplification:

Article Title: Translation of a Retained Intron in Tyrosinase-related Protein (TRP) 2 mRNA Generates a New Cytotoxic T Lymphocyte (CTL)-defined and Shared Human Melanoma Antigen Not Expressed in Normal Cells of the Melanocytic Lineage
Article Snippet: .. Expression of TRP-2 and Its Partially Spliced Form TRP-2–INT2. cDNA corresponding to 150 ng of total RNA from melanoma lines and fresh samples (tumors, skin, and retina) was amplified by PCR in 25 μl of water containing 200 μM of each dNTP, 0.6 μM of each primer, 1× PCR buffer, 1 U of Taq-Gold (all reagents from Perkin-Elmer Corp. ). .. TRP-2 cDNA was amplified using the sense primer PR3 located in exon 2 (5′-TTCGGCAGAACATCCATTCC-3′) and the TRP-2L antisense primer, originating an amplicon of 1186 bp.

Synthesized:

Article Title: Expression of a Truncated, Kinase-Defective TGF-? Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis
Article Snippet: .. For RT PCR analysis, cDNA was synthesized from 1 μg of total RNA pooled from the hind limbs of two to four mice using oligo dT primers as described in the GeneAmp RNA PCR kit (Perkin Elmer, Norwalk, CT). .. PCR amplification was performed using 5 μl of the cDNA mix.

Article Title: STRL33, A Novel Chemokine Receptor-like Protein, Functions as a Fusion Cofactor for Both Macrophage-tropic and T Cell Line-tropic HIV-1
Article Snippet: .. Amplifications were done with cDNA synthesized from 0.015 μg of poly(A)+ RNA, with 1.5 μM of each primer pool in a 20 μl reaction volume with Taq polymerase and reagents from Perkin Elmer (Norwalk, CT), according to the instructions of the manufacturer. .. PCR was done using 30 cycles of denaturation at 94°C for 0.5 min, annealing at 45°C for 2 min, and chain extension at 72°C for 1.5 min. 1 μl from the first PCR was used in a second PCR done identically to the first and the products of the second reaction were separated on a 1.5% agarose gel from which fragments of the predicted size of ∼670 bp were purified and inserted by blunt end ligation into the vector pNOTA/T7 (5′ 3′ Prime, Inc., Boulder, CO).

Isolation:

Article Title: TRAM2 Protein Interacts with Endoplasmic Reticulum Ca2+ Pump Serca2b and Is Necessary for Collagen Type I Synthesis
Article Snippet: .. Rat TRAM2 expression was estimated by RT-PCR of 100 ng of total RNA or 20 ng of poly(A)+ RNA, isolated from quiescent or activated HSCs, using a TthRT-PCR kit (Perkin-Elmer) in the presence of [32 P]dCTP, as described previously ( ). ..

Northern Blot:

Article Title: nc886 is induced by TGF-β and suppresses the microRNA pathway in ovarian cancer
Article Snippet: .. To detect nc886 (as well as vtRNA1-1 and 5S rRNA) by northern hybridization, RNA was resolved in a 15% denaturing polyacrylamide gel (with 7 M urea) and was transferred to Genescreen Plus Hybridization Transfer Membrane (Perkin-Elmer, Waltham, MA). .. The membrane was subjected to hybridization with 32 P-labeled probes (whose sequences are in Supplementary Data File ) in ULTRAhyb®-Oligo Buffer (Invitrogen), followed by wash with a buffer containing 2× SSC and 1% sodium dodecyl sulfate (SDS) for 5 min at room temperature (RT) twice and then 30 min at 37 °C.

Labeling:

Article Title: A Screen for RNA-Binding Proteins in Yeast Indicates Dual Functions for Many Enzymes
Article Snippet: .. RNA was fluorescently labeled with either Cy3 or Cy5 using the MICROMAX ASAP RNA labeling Kit (Perkin Elmer Cat# MPS544) according to the manufacturer's protocol. .. Labeled RNA was purified with the RNeasy Micro kit (Qiagen) to remove unincorporated dyes and immediately used for array analysis.

Mouse Assay:

Article Title: Expression of a Truncated, Kinase-Defective TGF-? Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis
Article Snippet: .. For RT PCR analysis, cDNA was synthesized from 1 μg of total RNA pooled from the hind limbs of two to four mice using oligo dT primers as described in the GeneAmp RNA PCR kit (Perkin Elmer, Norwalk, CT). .. PCR amplification was performed using 5 μl of the cDNA mix.

Reverse Transcription Polymerase Chain Reaction:

Article Title: TRAM2 Protein Interacts with Endoplasmic Reticulum Ca2+ Pump Serca2b and Is Necessary for Collagen Type I Synthesis
Article Snippet: .. Rat TRAM2 expression was estimated by RT-PCR of 100 ng of total RNA or 20 ng of poly(A)+ RNA, isolated from quiescent or activated HSCs, using a TthRT-PCR kit (Perkin-Elmer) in the presence of [32 P]dCTP, as described previously ( ). ..

Article Title: Expression of a Truncated, Kinase-Defective TGF-? Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis
Article Snippet: .. For RT PCR analysis, cDNA was synthesized from 1 μg of total RNA pooled from the hind limbs of two to four mice using oligo dT primers as described in the GeneAmp RNA PCR kit (Perkin Elmer, Norwalk, CT). .. PCR amplification was performed using 5 μl of the cDNA mix.

Article Title: Vitamin E inhibits CD95 ligand expression and protects T cells from activation-induced cell death
Article Snippet: .. One microgram of total RNA was reverse-transcribed using the RT-PCR kit (Perkin-Elmer Applied Biosystems, Foster City, California, USA). .. Aliquots were amplified in a DNA thermocycler (Stratagene, Heidelberg, Germany) with 2.5 U recombinant Taq polymerase (Sigma-Aldrich) as described previously ( ).

Expressing:

Article Title: TRAM2 Protein Interacts with Endoplasmic Reticulum Ca2+ Pump Serca2b and Is Necessary for Collagen Type I Synthesis
Article Snippet: .. Rat TRAM2 expression was estimated by RT-PCR of 100 ng of total RNA or 20 ng of poly(A)+ RNA, isolated from quiescent or activated HSCs, using a TthRT-PCR kit (Perkin-Elmer) in the presence of [32 P]dCTP, as described previously ( ). ..

Article Title: Translation of a Retained Intron in Tyrosinase-related Protein (TRP) 2 mRNA Generates a New Cytotoxic T Lymphocyte (CTL)-defined and Shared Human Melanoma Antigen Not Expressed in Normal Cells of the Melanocytic Lineage
Article Snippet: .. Expression of TRP-2 and Its Partially Spliced Form TRP-2–INT2. cDNA corresponding to 150 ng of total RNA from melanoma lines and fresh samples (tumors, skin, and retina) was amplified by PCR in 25 μl of water containing 200 μM of each dNTP, 0.6 μM of each primer, 1× PCR buffer, 1 U of Taq-Gold (all reagents from Perkin-Elmer Corp. ). .. TRP-2 cDNA was amplified using the sense primer PR3 located in exon 2 (5′-TTCGGCAGAACATCCATTCC-3′) and the TRP-2L antisense primer, originating an amplicon of 1186 bp.

Polymerase Chain Reaction:

Article Title: Expression of a Truncated, Kinase-Defective TGF-? Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis
Article Snippet: .. For RT PCR analysis, cDNA was synthesized from 1 μg of total RNA pooled from the hind limbs of two to four mice using oligo dT primers as described in the GeneAmp RNA PCR kit (Perkin Elmer, Norwalk, CT). .. PCR amplification was performed using 5 μl of the cDNA mix.

Article Title: Translation of a Retained Intron in Tyrosinase-related Protein (TRP) 2 mRNA Generates a New Cytotoxic T Lymphocyte (CTL)-defined and Shared Human Melanoma Antigen Not Expressed in Normal Cells of the Melanocytic Lineage
Article Snippet: .. Expression of TRP-2 and Its Partially Spliced Form TRP-2–INT2. cDNA corresponding to 150 ng of total RNA from melanoma lines and fresh samples (tumors, skin, and retina) was amplified by PCR in 25 μl of water containing 200 μM of each dNTP, 0.6 μM of each primer, 1× PCR buffer, 1 U of Taq-Gold (all reagents from Perkin-Elmer Corp. ). .. TRP-2 cDNA was amplified using the sense primer PR3 located in exon 2 (5′-TTCGGCAGAACATCCATTCC-3′) and the TRP-2L antisense primer, originating an amplicon of 1186 bp.

Hybridization:

Article Title: nc886 is induced by TGF-β and suppresses the microRNA pathway in ovarian cancer
Article Snippet: .. To detect nc886 (as well as vtRNA1-1 and 5S rRNA) by northern hybridization, RNA was resolved in a 15% denaturing polyacrylamide gel (with 7 M urea) and was transferred to Genescreen Plus Hybridization Transfer Membrane (Perkin-Elmer, Waltham, MA). .. The membrane was subjected to hybridization with 32 P-labeled probes (whose sequences are in Supplementary Data File ) in ULTRAhyb®-Oligo Buffer (Invitrogen), followed by wash with a buffer containing 2× SSC and 1% sodium dodecyl sulfate (SDS) for 5 min at room temperature (RT) twice and then 30 min at 37 °C.

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