(1 reference)Quick Overview for Nuclear/Cytosolic Fractionation Kit ()
应用范围
样品类型
-
-
Key Features
-
- Frequently cited in scientific publications with 300+ citations
- Validated inhouse for relevant applications
- Extensive validation report for IF provided by one of your peers
- Designed to detect RFP and its variants
-
Key Features
-
- Multiplex ELISA: Testing 5 Targets in 1 Run
- High performance COVID-19 antibody testing
- 99.6% specificity, 95.9% sensitivity
-
Key Features
-
- Recombinant human neutralizing antibody (hNAb) against SARS-CoV-2.
- Binds SARS-CoV-2 S proteins of lineages B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma), B.1.617.2 (delta), B.1.1.529 (omicron), B.1.429 (epsilon), B.1.525 (eta), and B.1.617.1 (kappa).
- Frequently used as reference in S-protein ELISAs and neutralization assays.
- Synergizes with other hNAbs: binds a highly conserved epitope, not interefering with the S-protein's ACE2 RBD.
-
Key Features
-
- Recombinant human neutralizing antibody (hNAb) against SARS-CoV-2.
- Binds SARS-CoV-2 S proteins of lineages B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma), B.1.429 (epsilon), and B.1.525 (eta).
- Frequently used as reference in S-protein ELISAs and neutralization assays.
- Synergizes with other hNAbs: binds a highly conserved epitope.
- No interference with the ACE2 binding site.
-
Key Features
-
- Dendra2 Antibody (ABIN361314) independently validated for Immunocytochemistry by Neurology Lab (INSERM).
- Publications cite use for Immunoblotting as well as Immunostaining.
- Dendra2 belongs to a new class of photoactivatable fluorescent proteins (PAFPs). PAFPs allow to photolable and track fusion proteins in vivo. Dendra2 Antibody (ABIN361314) can be used for control purposes in this setting.
-
Key Features
-
- Magnetic agarose concanavalin A beads for CUT&RUN and CUT&Tag assays.
- Superior handling compared to magnetic silica concanavalin A beads: easier washes and resuspension.
- Reduced risk for samples to dry out.
- Performance comparable to or better than magnetic silica concanavalin A beads.
-
Key Features
-
- Recombinant SARS-CoV-2 spike protein produced in HEK-293.
- Contains the E484K mutation (present in the novel lineage 501Y.S2 from South Africa) that can affect antibody recognition and enable SARS-CoV-2 immune escape.
- Contains the D614G mutation that renders SARS-CoV-2 infection and replication more efficient.
-
Key Features
-
- Recombinant SARS-CoV-2 spike protein RBD produced in HEK-293.
- Contains the spike protein N501Y mutation (present in the novel lineages B.1.1.7 and 501Y.S2) that has been associated with an increased transmission rate of SARS-CoV-2.
-
Key Features
-
- Recombinant SARS-CoV-2 spike protein RBD produced in HEK-293.
- Contains the spike protein N501Y mutation (present in the novel lineages B.1.1.7 and 501Y.S2) that has been associated with an increased transmission rate of SARS-CoV-2.
-
Key Features
-
- Simultaneous detection of Sp Cas9 binding and cleavage sites in genomic DNA.
- In situ assay leaves cells intact and reduces background.
- Unbiased detection of off-site effects because it is not based on a predictive algorithm.
-
Key Features
-
- Detection of SARS-CoV-2 with variety of different immunoassays, i.e. ELISA, Lateral Flow, as validated by numerous publications (see below).
- Nucleocapsid antibody used for diagnostic Covid19 testing as well as for drug discovery.
- Developed and manufactured in the U.S.
-
Key Features
-
- Has been shown to neutralize SARS-Cov-2 S protein RBD from lineages B.1.1.7, B.1.351, P.1, and B.1.429.
- Suitable as positive control in neutralization antibody screening.
- Humanized chimeric antibody consisting of human IgG1 constant domains and murine variable regions.
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of concern B.1.617.
- Contains E154K, L452R, E484Q, D614G, and P681R mutations characteristic for the SARS-CoV-2 kappa variant (B.1.617.1).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of interest B.1.525.
- Contains Q52R, del 69-70, del 144, E484K, F888L, and Q677H mutations characteristic for the SARS-CoV-2 eta variant (B.1.525).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of interest B.1.429.
- Contains S13I, W152C, L452R, and D1183Y mutations characteristic for the SARS-CoV-2 eta variant (B.1.429).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of concern B.1.1.7.
- Contains del 69-70, del 144, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H mutations characteristic for the SARS-CoV-2 alpha variant (B.1.1.7).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of concern P.1.
- Contains L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, H655Y, T1027I, and V1176F mutations characteristic for the SARS-CoV-2 gamma variant (P.1).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of concern B.1.617.
- Contains T19R, L452R, T478K, D614G, P681R and D950N mutations characteristic for the SARS-CoV-2 delta variant (B.1.617.2).
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of interest B.1.621 mu.
- Contains T95I, Y144S, Y145N, R346K, E484K, N501Y, D614G, P681H and D950N mutations characteristic for the SARS-CoV-2 mu variant (B.1.621).
-
Key Features
-
- NLRP3 antibody for reliable recognition of mouse and human NLRP3/NALP3
- This un-conjugated monoclonal antibody has 60 PubMed citations
-
Key Features
-
- 95+ publication references, 1 independent validation
- Frequently used as CUT&RUN IgG negative control and CUT&Tag secondary antibody
- Used in CUT&RUN and CUT&Tag protocols, e.g. Henikoff et al. (2018) PMID 29651053, Kaya-Okur et al. (2019, 2020) PMID 31036827 and PMID 32913232
-
Key Features
-
- Recombinant trimeric S protein of the SARS-CoV-2 variant of concern B.1.1.529 omicron (21K).
- Contains A67V, H69del, V70del, T95I, G142del, V143del, Y144del, Y145D, N211del, L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, and L981F mutations characteristic for the SARS-CoV-2 omicron variant (B.1.1.529).
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern B.1.617.2 delta expressed in insect cells.
- Contains T19R, G142D, E156G, FR157-158 deletion, L452R, T478K, D614G, P681R, and D950N mutations characteristic for the SARS-CoV-2 delta variant (B.1.617.2).
- Intact furin cleavage site 680-SRRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern B.1.617.2 delta expressed in insect cells.
- Contains EFR157-158 deletion, T19R, G142D, E156G, FR157-158 deletion, L452R, T478K, D614G, P681R, and D950N mutations characteristic for the SARS-CoV-2 delta variant (B.1.617.2).
- Intact furin cleavage site 680-SRRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern P.1 gamma expressed in insect cells.
- Contains L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, and T1027I mutations characteristic for the SARS-CoV-2 gamma variant (P.1).
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern P.1 gamma expressed in insect cells.
- Contains L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, and V1176F mutations characteristic for the SARS-CoV-2 gamma variant (P.1).
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 (M1-P1213) of the SARS-CoV-2 variant of concern B.1.351 expressed in insect cells.
- Contains D80A, K417N, E484K, N501Y, D614G, and A701V mutations characteristic for the SARS-CoV-2 beta variant (B.1.351).
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern B.1.351 expressed in insect cells.
- Contains L18F, D80A, D215G, LAL242-244 deletion, R246I, K417N, E484K, N501Y, D614G, and A701V mutations characteristic for the SARS-CoV-2 beta variant (B.1.351).
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 (M1-P1213) of the SARS-CoV-2 variant of concern B.1.351 expressed in insect cells.
- Contains L18F, D80A, D215G, LAL242-244 deletion, R246I, K417N, E484K, N501Y, D614G, and A701V mutations characteristic for the SARS-CoV-2 beta variant (B.1.351).
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern B.1.1.7 expressed in insect cells.
- Contains HV69-70 deletion, Y145 deletion, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H characteristic for the SARS-CoV-2 alpha variant (B.1.1.7).
- Intact furin cleavage site 680-SHRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant S protein S1+S2 ECD (M1-P1213) of the SARS-CoV-2 variant of concern B.1.1.7 expressed in insect cells.
- Contains HV69-70 deletion, Y144 deletion, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H characteristic for the SARS-CoV-2 alpha variant (B.1.1.7).
- Intact furin cleavage site 680-SHRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- Recombinant SARS-CoV-2 wt S protein S1+S2 (M1-P1213) expressed in insect cells.
- Contains D614G mutation.
- Intact furin cleavage site 680-SPRRAR|SV-687.
- C-terminal His-tag.
-
Key Features
-
- High-affinity single-domain antibody
- Recognizes mRFP, mCherry, mScarlet-i, tdTomato, dsRed etc.
- Bead size 50-150 μm
-
Key Features
-
- Monoclonal murine neutralizing antibody (nAb) against SARS-CoV-2 S protein.
- Binds SARS-CoV-2 S proteins of various lineages, including B.1.617.2 (delta) and B.1.1.529 (omicron).
- Ideal for SARS-CoV-2 S-protein ELISAs and neutralization assays.
-
Key Features
-
- Monoclonal murine neutralizing antibody (nAb) against SARS-CoV-2 S protein.
- Binds SARS-CoV-2 S proteins of various lineages, including B.1.617.2 (delta). Does NOT bind the S protein of SARS-CoV-2 B.1.1.529 (omicron).
- Ideal for SARS-CoV-2 S-protein ELISAs and neutralization assays.
-
Key Features
-
- Rabbit beta-Catenin antibody reliably detects beta-Catenin targets in Wnt-signaling using CUT&RUN, ICC, IF, WB, ELISA.
- Well suited to image adherens junctions in IHC/IF.
- Highly specific beta-Catenin detection in WB.
-
Key Features
-
- Rabbit anti-CRBN Antibody reliably detects CRBN in human species.
- Polyclonal, unconjugated antibody for ELISA, WB, IHC.
- Highly specific CRBN detection. See validation images.
-
Key Features
-
- Mouse IL-1 beta ELISA kit for quantitative detection of IL-1 beta.
- Reliable product with validated components.
- ELISA kit cited in more than 40 PubMed References.
-
Key Features
-
- Magnetic agarose Concanavalin A beads for affinity chromatography using a magnetic separator.
- Easy bench-top purification of viruses, virus-like particles (VLP), and cultured cells via reversible binding of glycoproteins, glycolipids, or polysaccharides with terminal mannose or glucose to concanavalin A.
-
Key Features
-
- Polyclonal, unconjugated GFP antibody for reliable detection of GFP and its variants.
- Validated for Fluorescence Microscopy, ELISA, Western Blotting
- High Quality GFP antibody, cited in more than 177 PubMed References.
- Available in 10 µl and 100 µl quantities.
-
Key Features
-
- Reliable Taurine Assay Kit for measurement of free taurine in biological samples.
- Sample taurine concentrations are determined by comparison with a known taurine standard.
- Quick and easy to use.
-
Key Features
-
- Impurity detection in the manufacturing of biological drugs.
- The AccuSignal™ Nuclease ELISA Kit is designed for sensitive and reliable quantification of nucleases in therapeutic products, including DENARASE®, Benzonase®, and Turbonuclease.
- Offers a broad quantification spectrum, maintaining outstanding dilution linearity, instilling trust in the accuracy of results over a wide array of nuclease concentrations.
- Delivers consistent and repeatable outcomes, characterized by minimal intra- and inter-assay variability.
- Tailored antibody specificity allows for application across diverse materials, accommodating various nuclease products from multiple suppliers.
- Components: 96-well strip plate, plate sealer, nuclease standard, biotinylated detection antibody, Streptavidin-HRO (100X), buffers.
-
Key Features
-
- High-affinity anti-GFP single-domain antibody (sdAb) covalently bound to 4 % cross-linked agarose magnetic beads with 50-150 µm diameter.
- The Alpaca sdAb clone 1H1 is specific for GFP and many GFP derivatives. Capacity > 3 μg GFP per μl of packed beads.
- Compatible with physiological buffers, common Lysis and Wash Buffers, and high stringency buffers.
-
Key Features
-
- High-quality FcRn antibody (Clone ADM31) for the detection of FcRn.
- Highly specific for human FcRn, blocks the binding of human serum albumin.
- PBS-only, preservative free
- 15 PubMed references available for this FcRn antibody.
-
Key Features
-
- High-quality FcRn antibody (Clone DVN24) for the detection of FcRn.
- Highly specific for human FcRn, blocks the binding of IgG.
- PBS-only, preservative free
- 2 PubMed references available for this FcRn antibody.
-
Key Features
-
- High-affinity anti-GFP single-domain antibody (sdAb) covalently bound to 4 % cross-linked agarose beads with 50-150 µm diameter.
- The Alpaca sdAb clone 1H1 is specific for GFP and many GFP derivatives. Capacity > 3 μg GFP per μl of packed beads.
- Compatible with physiological buffers, common Lysis and Wash Buffers, and high stringency buffers.
-
Key Features
-
- TrueBlot®: Anti-Rabbit IgG HRP, Mouse Monoclonal eB182
- Manufactured by Rockland Immunochemicals, Inc.
- Rockland Product ID: 18-8816-33
-
Key Features
-
- TrueBlot®: Anti-Rabbit IgG HRP, Mouse Monoclonal eB182
- Manufactured by Rockland Immunochemicals, Inc.
- Rockland Product ID: 18-8816-31
-
Key Features
-
- Anti-ATM Protein Kinase pS1981 (MOUSE) Monoclonal Antibody
- Manufactured by Rockland Immunochemicals, Inc.
- Rockland Product ID: 200-301-400
-
Key Features
-
- High-quality FcRn antibody (Clone ADM31) for the detection of FcRn.
- Highly specific for human FcRn, blocks the binding of human serum albumin.
- contains Glycerol for better storage.
-
Key Features
-
- High-quality FcRn antibody (Clone DVN24) for the detection of FcRn.
- Highly specific for human FcRn, blocks the binding of IgG.
- Contains Glycerol for better storage.
-
产品特性
- Nuclear/Cytosolic Fractionation Kit provides a simple and fast tool to isolate nuclear extract from the cytoplasmic fraction of mammalian cells. The procedure has been optimized to provide extraction, with high protein recovery and low cross-contamination, in less than 2 hours. The extracted protein fractions are functional and suitable for downstream assays such as DNA footprinting, RNA splicing, gel shift assays (EMSA), reporter assays, enzyme activity assays, and 6 Western blotting. Each kit provides sufficient quantities to perform 20 preps (up to 5 x 10 cells each).
-
组件
-
- Cytosol Extraction Buffer, Hypotonic (10X) : One vial - 2 mL.
- Cell Lysis Reagent : One vial - 1 mL of 10% Igepal CA-630 in 1X Cytosol Extraction Buffer (CEB).
- Nuclear Extraction Buffer : One vial - 2 mL.
- Dithiothreitol (1000X) : One vial - 20 μL of 1 M DTT.
- Protease Inhibitor Cocktail (100X) : One vial - 200 μL containing AEBSF, Aprotinin, Bestatin, E64, Leupeptin, and Pepstatin A in DMSO.
-
试剂未包括
-
- PBS
- Microcentrifuge tubes
- Microcentrifuge
-
Product Specific Information
-
Our GFP Catcher is a product based on the GFP pull-down technique used to isolate GFP fusion proteins from cell lysates. It consists of special magnetic beads (product ABIN7272855) or agarose beads (product ABIN5311508) coated with an antibody against GFP. These beads allow efficient and selective binding to GFP or GFP-fused target proteins.
The application of GFP Catcher is similar to the GFP pull-down method. First, the protein of interest is expressed in the cells with it fused to GFP. Then, the cell lysate is prepared to extract the target proteins. The GFP catcher beads are then added to the cell lysate and bound to these proteins by specifically binding to the GFP or GFP fusion protein.
After the beads with the bound target proteins are isolated, thorough purification is performed to remove components that are not specifically bound. Finally, the target proteins can be separated from the beads and used for further analysis such as immunoblotting or mass spectrometry.
GFP Catcher offers the advantage of quick and easy isolation of GFP fusion proteins. It is a useful tool for protein analysis and allows efficient identification of proteins interacting with GFP fusion protein. GFP Catcher is widely used in cell biology research to study protein-protein interactions and deepen the understanding of cellular processes.
-
-
Tumor Necrosis Factor alpha (TNF alpha) ELISA Kit
TNF alpha 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Interleukin 1, beta (IL1B) ELISA KitIL1B 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 小鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 人 Colorimetric Sandwich ELISA 62.5 pg/mL - 4000 pg/mL Plasma, Serum
Amyloid beta 1-40 (Abeta 1-40) ELISA KitAbeta 1-40 适用: 人 Colorimetric Sandwich ELISA 7.813 pg/mL - 500 pg/mL Plasma, Serum, Tissue Homogenate
Amyloid beta 1-42 (Abeta 1-42) ELISA KitAbeta 1-42 适用: 人 Colorimetric Sandwich ELISA 4.688 pg/mL - 300 pg/mL Plasma, Serum, Tissue Homogenate
Lipopolysaccharides (LPS) ELISA Kit适用: Various Species Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 小鼠 Colorimetric Sandwich ELISA 46.88 pg/mL - 3000 pg/mL Plasma, Serum
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 人 AA 77-233 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
Interleukin 6 (IL6) ELISA KitIL6 适用: 人 Colorimetric Sandwich ELISA 7.8-500 pg/mL Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate, Urine
-
-
应用备注
- Optimal working dilution should be determined by the investigator.
-
说明
-
- Simple and fast tool to isolate nuclear extract from the cytoplasmic fraction of mammalian cells
- Extraction in less than 2 hours
- Each prep can accommodate up to 5 x 106cells
-
试剂准备
-
1X Cytosol Extraction Buffer (CEB): Dilute the 10X Cytosol Extraction Buffer to 1X with deionized water. Stir to homogeneity. Dithiothreitol: Immediately before use dilute the Dithiothreitol 1:1000 with 1X Cytosol or Nuclear Extraction Buffer. Stir to homogeneity. Do not store diluted solutions. 2 Protease Inhibitor Cocktail: Immediately before use dilute the Protease Inhibitor Cocktail 1:100 with 1X Cytosol or Nuclear Extraction Buffer. Stir to homogeneity. Do not store diluted solutions.
-
样品制备
-
I. Adherent Cells
- Culture cells to approximately 80-90 % confluence.
- Aspirate the culture media and wash twice with PBS.
- Detach the cells from the plates in PBS by scraping with a cell scraper.
- Collect the solution into an appropriate conical centrifuge tube.
- Centrifuge for 5 minutes (600 x g).
- Discard the supernatant and immediately proceed to the Assay Protocol.
II. Suspension Cells
- Collect the cells into an appropriate conical centrifuge tube.
- Centrifuge for 5 minutes (600 x g).
- Remove and discard the supernatant.
- Wash the cells twice with PBS.
- Centrifuge for 5 minutes at (600 x g).
- Discard the supernatant and immediately proceed to the Assay Protocol.
-
实验流程
-
Important Note: Perform the below steps at 2-8 °C. All buffers, centrifuge rotors, and equipment should be maintained at 2-8 °C. Before use, Dithiothreitol and Protease Inhibitor Cocktail should be diluted according to the Preparation of Reagents section above.
I. Cytosol Fractionation Protocol
- Collect cells (up to 5 x 10 ) by centrifugation for 5 minutes at 4 °C (600 x g).
- Wash the cells once with ice cold PBS.
- Remove and discard the supernatant.
- Gently resuspend the cell pellet with 500 μL of ice cold, 1X Cytosol Extraction Buffer (containing DTT/Protease Inhibitors) by pipetting up and down.
- Transfer the suspension into a prechilled microcentrifuge tube.
- Incubate on ice for 10 minutes.
- Add 25 μL of Cell Lysis Reagent and vortex for 10 seconds at the highest setting. 3
- Centrifuge for 10 minutes at 4 °C (800 x g).
- Carefully transfer the supernatant (cytoplasmic fraction) to a clean, chilled microcentrifuge tube. The cytoplasmic fraction can be stored at -80 °C for future use. Note: Make sure not to disturb/remove the nuclei pellet.
- Gently resuspend the pellet with 500 μL of ice cold, 1X Cytosol Extraction Buffer (containing DTT/Protease Inhibitors) by pipetting up and down. Note: This wash step is included to reduce cross-contamination between fractions.
- Add 25 μL of Cell Lysis Reagent and vortex for 10 seconds at the highest setting.
- Centrifuge for 10 minutes at 4 °C (800 x g).
- Carefully aspirate the supernatant and discard of this wash. II. Nuclear Protein Extraction Protocol
- Gently resuspend the nuclear pellet with 100 μL of ice cold, 1X Nuclear Extraction Buffer (containing DTT/Protease Inhibitors) by pipetting up and down.
- Maintain on ice for 30 minutes, vortexing for 10 seconds at the highest setting in 10 minute intervals.
- Centrifuge for 30 minutes at 4 °C (14000 x g).
- Carefully transfer the supernatant (nuclear protein extract) to a clean, chilled microcentrifuge tube. The extract can be stored at -80 °C for future use. Note: The nuclear extract typically yields protein concentrations of > 1 mg/mL. If greater concentrations are desired, resuspend the nuclear pellet in a smaller volume in step 1 above (minimum of 25 μL).
III. Other Considerations For determining the protein content of extracts, samples must be diluted 1:2 before running in the Bradford Protein Assay. Buffer only controls must be performed concurrently. DTT in the buffers is not compatible with the BCA Protein Assay. Nuclear Extraction Buffer is a high salt buffer, containing 420 mM NaCl. If salt removal is necessary, dialysis or a desalting column may be used. 4 Example of Results The following figure demonstrates typical results seen with Cell Biolabs' Nuclear/Cytosolic Fractionation Kit. One should use the data below for reference only. Figure 1: HEK293 Cell Fractionation. Cytosolic and nuclear protein extracts were isolated from Human Embryonic Kidney 293 cells according to the Assay Protocol. Whole cell (W), cytosol (C), and nuclear (N) fractions were immunoblotted with Anti-α-Tubulin (left) or Anti-Lamin A/C (right) at 1 μg/mL. Note: Anti-α-Tubulin (Calbiochem CP06) and Anti-Lamin A/C (Sigma SAB4200236) are both mouse monoclonals. Tubulin and Lamin are known to be cytosolic and nuclear specific proteins, respectively. Figure 2: HEK293 Trypan Blue Staining. Human Embryonic Kidney 293 cells were stained with Trypan Blue at various steps during the fractionation protocol, demonstrating complete lysis and high neuclei recovery. Recent Product Citations
- Davis, M. R. et al. (2016). Epigenetically maintained SW13+ and SW13-subtypes have different oncogenic potential and convert with HDAC1 inhibition. BMC Cancer. doi:10.1186/s12885-016- 2353-7. 5
- Zhang, P. et al. (2016). An oxygen-insensitive Hif-3a isoform inhibits Wnt signaling by destabilizing the nuclear b-catenin complex. eLife. doi:10.7554/eLife.08996.
- Nakamura, S. et al. (2015). Novel roles for LIX1L in promoting cancer cell proliferation through ROS1-mediated LIX1L phosphorylation. Sci Rep. doi:10.1038/srep13474.
- Shinmura, K. et al. (2015). NEIL1 p. Gln282Stop variant is predominantly localized in the cytoplasm and exhibits reduced activity in suppressing mutations. Gene. doi:10.1016/j.gene.2015.06.043.
- Jeon, Y. J. et al. (2015). A set of NF-κB-regulated microRNAs induces acquired TRAIL resistance in lung cancer. Proc Natl Acad Sci U S A. 112:E3355-64.
- Ohtsuka, S. et al. (2014). SQSTM1/p62/A170 regulates the severity of Legionella pneumophila pneumonia by modulating inflammasome activity. Eur J Immunol. 44:1084-1092.
- Zou, J. et al. (2014). A TIR domain protein from E. faecalis attenuates MyD88-mediated signaling and NF-κB activation. PLoS One. 9:e112010.
-
限制
- 仅限研究用
-
-
-
注意事项
- Avoid multiple freeze/thaw cycles.
-
储存条件
- 4 °C/-20 °C
-
储存方法
- Upon receiving, aliquot and store Dithiothreitol and Protease Inhibitor Cocktail at -20°C and avoid multiple freeze/thaw cycles. Store all other components at 4°C.
-
-
-
: "The downregulation of GFI1 by the EZH2-NDY1/KDM2B-JARID2 axis and by human cytomegalovirus (HCMV) associated factors allows the activation of the HCMV major IE promoter and the transition to ..." in: PLoS pathogens, Vol. 10, Issue 5, pp. e1004136, (2014) (PubMed).
-
: "The downregulation of GFI1 by the EZH2-NDY1/KDM2B-JARID2 axis and by human cytomegalovirus (HCMV) associated factors allows the activation of the HCMV major IE promoter and the transition to ..." in: PLoS pathogens, Vol. 10, Issue 5, pp. e1004136, (2014) (PubMed).
-
