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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Immunocytochemistry [1]
- Flow cytometry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- 701778 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Acetyl-Histone H4 (Lys20) Antibody (2H2L2), 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
- 2H2L2
- 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
- Immunofluorescence was performed on fixed and permeabilized MCF7 cells for detection of Histone H4K20ac using ABfinity Anti-Histone H4K20ac Recombinant Rabbit Monoclonal Antibody (Product # 701778, 1 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of Histone H4K20ac protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Histone H4K20ac. Panel e) represents control cells with no primary antibody to assess background.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry analysis of Histone H4K20ac was performed on Jurkat cells labeled with ABfinityª Anti-Histone H4K20ac Recombinant Rabbit Monoclonal Antibody (Product# 701778, 2-4 ug/ 1M cells) or with rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonalª 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 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
- Enrichment of endogenous Acetyl-Histone H4 (Lys20) protein at specific gene loci using ABfinity™ Anti-Acetyl-Histone H4 (Lys20) Rabbit Monoclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using ABfinity™ Anti-Acetyl-Histone H4 (Lys20) Rabbit Monoclonal Antibody (Product # 701778, 3 ug) 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 qPCR with PCR primer pairs for the promoters of CCND2, C-FOS as positive, and SAT2 satellite repeats used as negative target genes/binding sites. 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-Acetyl-Histone H4 (Lys20) Oligoclonal Antibody (Product # 701778, 1:2000 dilution) to Histone H4K20ac 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).
