S100A13 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Protein S100-A13, S100 calcium-binding protein A13, S100a13, S10AD_HUMAN
S100A13 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
S100 Calcium Binding Protein A13 also known as S100A13 is a member of the S100 protein family and displays calcium-binding abilities usually binding two calcium ions. This small protein has a molecular mass of approximately 11 kDa. S100A13 is expressed in a variety of tissues including the heart brain and skeletal muscle. Its expression can be altered in different cellular conditions suggesting an adaptive role in diverse physiological contexts.
S100A13 influences several cellular processes related to growth and differentiation. This protein facilitates secretion of fibroblast growth factor 1 (FGF1) and interleukin-1 alpha (IL-1α) through a non-classical pathway. S100A13 does not function alone; it engages in complexes with other proteins to execute its roles. It often works together with S100A6 and calcyclin-binding protein in these secretory pathways.
S100A13 participates in the regulation of angiogenesis and inflammation. It is involved in the FGF signal transduction pathway by affecting the externalization of FGF1 an important player in angiogenesis. Additionally S100A13 contributes to inflammation-related processes. Its activity is linked to proteins such as p53 and FGF1 integrating it into wider cellular systems.
S100A13 is implicated in cancer and chronic inflammatory conditions. Overexpression of S100A13 has links to tumor progression and aggressiveness suggesting a role in cancer. In chronic inflammation S100A13 associates with increased levels of inflammatory cytokines. It connects to proteins like FGF1 and IL-1α in these contexts linking it with pathophysiological mechanisms that underlie these conditions.
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Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] ab109252 was shown to react with S100 Calcium Binding Protein A13/S100A13 in wild-type HEK-293T cells in Western blot with loss of signal observed in S100A13 knockout cell line ab266065 (S100A13 knockout cell lysate ab258183). Wild-type HEK-293T and S100A13 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] ab109252 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] ab109252) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: S100A13 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human S100A13 (S100 Calcium Binding Protein A13) knockout HEK-293T cell line (ab266065)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 13 kDa
Homozygous: Insertion of the selection cassette in exon 4
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