Human BRPF1 (Peregrin) knockout HEK-293T cell lysate (ab258795)
Overview
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Product name
Human BRPF1 (Peregrin) knockout HEK-293T cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 34 bp deletion in exon 2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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Tested applications
Suitable for: Sanger Sequencing, WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab261349 - Human BRPF1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Target
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Function
Component of the MOZ/MORF complex which has a histone H3 acetyltransferase activity. Positively regulates the transcription of RUNX1 and RUNX2. -
Tissue specificity
High levels in testis. -
Sequence similarities
Contains 1 bromo domain.
Contains 1 C2H2-type zinc finger.
Contains 1 PHD-type zinc finger.
Contains 1 PWWP domain. -
Post-translational
modificationsAcetylated by KAT6A. -
Cellular localization
Nucleus. Cytoplasm. Localization to the nucleus depends on KAT6A, ING5 and MEAF6. - Information by UniProt
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Alternative names
- BR140
- Bromodomain and PHD finger containing 1
- Bromodomain and PHD finger-containing protein 1
see all
Associated products
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KO cell lines
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab258795 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sanger Sequencing |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 137 kDa.
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| Notes |
|---|
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Sanger Sequencing
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 137 kDa. |
Images
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Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell lysate (ab258795)
Lane 1: Wild-type HEK-293T cell lysate 20 μg
Lane 2: BRPF1 knockout HEK-293T cell lysate 20 μg
Lane 3: BRPF1 knockout HEK-293T cell lysate 20 μg
Lane 4: Empty
Lane 5: PC-3 cell lysate 20 μg
Lane 6: HepG2 cell lysate 20 μg
False colour image of Western blot: Anti-Peregrin/BRPF1 antibody [EPR24069-57] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab259840 was shown to bind specifically to Peregrin/BRPF1. A band was observed at 120 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BRPF1 knockout cell line ab266549 (knockout cell lysate ab258795). To generate this image, wild-type and BRPF1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. -
Human BRPF1 (Peregrin) knockout HEK-293T cell lysate (ab258795)Lane 1: Wild-type HEK-293T cell lysate 20 μg
Lane 2: BRPF1 knockout HEK-293T cell lysate 20 μg
Lane 3: PC-3 cell lysate 20 μg
Lane 4: HepG2 cell lysate 20 μgFalse colour image of Western blot: Anti-Peregrin/BRPF1 antibody - N-terminal staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab226909 was shown to bind specifically to Peregrin/BRPF1. A band was observed at 160, 145 and 138 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BRPF1 knockout cell line ab266549 (knockout cell lysate ab258795). To generate this image, wild-type and BRPF1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Observed bands - 160, 145 and 138 kDa
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Sanger Sequencing - Human BRPF1 knockout HEK293T cell lysate (ab258795)Homozygous: 34 bp deletion in exon 2
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab258795 has not yet been referenced specifically in any publications.