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Western blot
Flow cytometryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Flow cytometry [1]
- Protein array [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- 701784 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Tri-Methyl-Histone H3 (Lys9) Antibody (21H2L16), ABfinity™ Rabbit Monoclonal
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- ABfinity recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A. ABfinity monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance. Since it is highly conserved across species, the antibody may react with many other species.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 21H2L16
- Vial size
- 100 µg
- Concentration
- 0.5 mg/ml
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry analysis of Histone H3K9me3 was performed on Jurkat labeled with ABfinity™ Anti-H3K9me3 Recombinant Rabbit Monoclonal Antibody (Product# 701784, 2-4 ug/ 1M cells) or with rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, (Alexa Fluor® 488 conjugate, Product # A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer (4468770).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- The specificity of the Antibody, ABfinity™ Anti-Histone H3K9me3 Recombinant Rabbit Monoclonal Antibody (Product # 701784, 1-2 ug/ml) to Histone H3K9me3 peptide was confirmed using MODified™ Histone Peptide Array (Product #13001, Active Motif). Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution) was used as secondary Antibody. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). The data was analyzed using Array analyze software as per manufacturer instructions.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) was performed using ABfinity™ Anti-Histone H3K9me3 Recombinant Rabbit Monoclonal Antibody (Product # 701784, 5ug) on sheared chromatin from 2 million HeLa cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the inactive SAT2 satellite repeat, used as positive control target gene, and promoters of the active cFOS, beta-ACTIN used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- The specificity of the antibody, ABfinity™ Anti-Tri-Methyl-Histone H3 (Lys9) Monoclonal Antibody (Product # 701784, 0.25 ug/ml) to Histone H3K9me3 peptide was confirmed using MODified™ Histone Peptide Array (Active Motif, 13001). Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) was used as the secondary antibody. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). The dot blot data obtained (lower panel) was analyzed using Array Analyze software as per the manufacturer's instructions (top panel).
