SIRPA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9,
Homozygous: 35 bp deletion in exon 6.
Images
Key facts
- Cell type
RAW 264.7
- Species or organism
Mouse
- Tissue
Lymphatic
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9,
Homozygous: 35 bp deletion in exon 6
Alternative names
Bit, Brain Ig-like molecule with tyrosine-based activation motifs, Brain immunoglobulin like molecule with tyrosine based activation motifs, CD172 antigen-like family member A, CD172a, CD172a antigen, Inhibitory receptor SHPS-1, MFR, MYD 1, Macrophage fusion receptor, MyD-1 antigen, PTPNS1, Protein tyrosine phosphatase non receptor type substrate 1, SHP substrate 1, SHPS1_HUMAN, SIRP, SIRPA, SIRPalpha, Signal regulatory protein alpha, Signal regulatory protein alpha type 1, Signal regulatory protein alpha type 2, Signal-regulatory protein alpha-1, Signal-regulatory protein alpha-2, Signal-regulatory protein alpha-3, Sirp-alpha-1, Sirp-alpha-2, Sirp-alpha-3, Tyrosine phosphatase SHP substrate 1, Tyrosine-protein phosphatase non-receptor type substrate 1, p84
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SIRPA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9,
Homozygous: 35 bp deletion in exon 6.
Key facts
- Cell type
RAW 264.7
- Species or organism
Mouse
- Tissue
Lymphatic
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9,
Homozygous: 35 bp deletion in exon 6- Disease
Carcinoma
- Concentration
Properties
- Gene name
SIRPA
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
- Zygosity
Homozygous
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Male
Storage
- Shipped at conditions
Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
SIRP alpha also known as CD172a is a transmembrane receptor protein with a mass ranging between 70-110 kDa due to glycosylation. This protein extensively expresses on the surface of myeloid cells neurons and a subset of T-cells and is part of the immunoglobulin superfamily. SIRP alpha interacts with its ligand CD47 a widely expressed glycoprotein involved in immune response regulation. Its mechanical action primarily involves signal regulation through the recruitment of SHP-1 and SHP-2 two cytoplasmic tyrosine phosphatases.
- Biological function summary
SIRP alpha functions significantly in the regulation of phagocytosis acting as a "don't eat me" signal to macrophages upon binding with CD47. It does not act alone; rather it is part of a complex that recruits SHP-1 and SHP-2 leading to inhibition of dephosphorylation activities essential for engulfment processes. This regulatory mechanism is important for maintaining cellular homeostasis ensuring that healthy cells are not mistakenly destroyed by the immune system.
- Pathways
SIRP alpha plays an important role in the innate immune pathways involving the regulation of phagocytosis and cell-cell adhesion. Particularly it fits into the immune checkpoint pathways where it interacts closely with proteins like CD47 and plays a role in the interaction between the immune system and cancer cells. Through these pathways SIRP alpha helps maintain balance in the immune response allowing for the recognition of self versus non-self therefore preventing autoimmunity while facilitating the clearance of pathogens.
- Associated diseases and disorders
SIRP alpha is implicated in cancer and autoimmune diseases. In cancer its interaction with CD47 allows tumor cells to escape phagocytosis promoting tumor survival and growth. In autoimmune disorders dysregulated SIRP alpha expression or signaling could miscommunicate immune signals leading to the destruction of healthy tissue. Understanding the link between SIRP alpha and these conditions can reveal potential targets for therapeutic development especially using inhibitors or modulators targeting the SIRP alpha-CD47 interaction.
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3 product images
Western blot - Mouse SIRPA knockout RAW 264.7 cell lysate (ab282969)
Lane 1: Wild-type RAW 264.7 cell lysate 20 μg
Lane 2: RAW 264.7 cell lysate 20 μg
Lane 3: THP-1 cell lysate 20 μg
Lane 4: MCF7 cell lysate 20 μg
False colour image of Western blot: Anti-SIRPA antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to SIRPA. A band was observed at 100-140 kDa (mouse SIRPA, isoform 1), in wild-type RAW 264.7 cell lysates (band observed at 70-100 kDa in THP-1 is Human SIRPA) with no signal observed at this size in SIRPA knockout cell line Mouse SIRPA knockout RAW 264.7 cell line ab281618 (knockout cell lysate ab282969). To generate this image, wild-type and SIRPA knockout RAW 264.7 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 (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) at 1/20000 dilution.All lanes: Western blot - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) at 1/20000 dilution
Lane 1: Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2: RAW 264.7 cell lysate at 20 µg
Lane 3: THP-1 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Predicted band size: 55 kDa
Western blot - Mouse SIRPA knockout RAW 264.7 cell lysate (ab282969)
Lane 1: Wild-type RAW 264.7 cell lysate 20 μg
Lane 2: SIRPA knockout RAW 264.7 cell lysate 20 μg
Lane 3: THP-1 cell lysate 20 μg
Lane 4: MCF7 cell lysate 20 μg
False colour image of Western blot: Anti-SIRP alpha antibody [EPR16264] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-SIRP alpha antibody [EPR16264] ab191419 was shown to bind specifically to SIRP alpha. A band was observed at 100-140 kDa (mouse SIRPA, isoform 1) & 40-50 kDa (mouse SIRPA, isoform 2), in wild-type RAW 264.7 cell lysates (band observed at 70-100 kDa in THP-1 is Human SIRPA) with no signal observed at this size in SIRPA knockout cell line Mouse SIRPA knockout RAW 264.7 cell line ab281618 (knockout cell lysate ab282969). To generate this image, wild-type and SIRPA knockout RAW 264.7 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 (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) at 1/20000 dilution.All lanes: Western blot - Anti-SIRP alpha antibody [EPR16264] (Anti-SIRP alpha antibody [EPR16264] ab191419) at 1/1000 dilution
Lane 1: Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2: SIRPA knockout RAW 264.7 cell lysate at 20 µg
Lane 3: THP-1 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 100-140 kDa
Sanger Sequencing - Mouse SIRPA knockout RAW 264.7 cell lysate (ab282969)
35 bp deletion in exon 6
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