Human HMGB1 knockout HeLa cell lysate
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(0 Publication)
- WB
Lab
Western blot - Human HMGB1 knockout HeLa cell lysate (AB263782)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : HMGB1 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab79823 observed at 30 kDa. Red - loading control ab8245 observed at 37 kDa.
ab79823 Recombinant Anti-HMGB1 antibody [EPR3507] was shown to specifically react with HMGB1 in wild-type HeLa cells in western blot. Loss of signal at the expected size was observed when knockout cell line ab255395 (knockout cell lysate ab263782) was used. The band observed in lane 2 below 25kDa may represent truncated forms and cleaved fragments. Wild-type and HMGB1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab79823 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-HMGB1 antibody [EPR3507] (<a href='/en-us/products/primary-antibodies/hmgb1-antibody-epr3507-ab79823'>ab79823</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HMGB1 knockout HeLa cell lysate at 20 µg
Predicted band size: 24 kDa
Observed band size: 30 kDa
false
- WB
Lab
Western blot - Human HMGB1 knockout HeLa cell lysate (AB263782)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : HMGB1 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab92310 observed at 30 kDa. Red - loading control ab8245 observed at 37 kDa.
ab92310 Recombinant Anti-HMGB1 antibody [EPR3506] was shown to specifically react with HMGB1 in wild-type HeLa cells in western blot. Loss of signal at the expected size was observed when knockout cell line ab255395 (knockout cell lysate ab263782) was used. The band observed in lane 2 below 25kDa may represent truncated forms and cleaved fragments. Wild-type and HMGB1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab92310 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-HMGB1 antibody [EPR3506] (<a href='/en-us/products/primary-antibodies/hmgb1-antibody-epr3506-ab92310'>ab92310</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HMGB1 knockout HeLa cell lysate at 20 µg
Predicted band size: 24 kDa
Observed band size: 30 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HMGB1 knockout HeLa cell lysate (AB263782)
Homozygous : 1 bp deletion in exon2
Complementary Products
Reactivity data
Product details
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
REACH authorisation
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.
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. 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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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