Mouse SIRPA knockout RAW 264.7 cell lysate
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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.
View 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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-SIRP alpha antibody [EPR16264] (<a href='/en-us/products/primary-antibodies/sirp-alpha-antibody-epr16264-ab191419'>ab191419</a>) 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 2:
Western blot - Mouse SIRPA knockout RAW 264.7 cell line (<a href='/en-us/products/cell-lines/mouse-sirpa-knockout-raw-2647-cell-line-ab281618'>ab281618</a>)
Lane 3:
THP-1 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 55 kDa
Observed band size: 100-140 kDa
false
- WB
Lab
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] (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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (<a href='/en-us/products/primary-antibodies/gapdh-antibody-6c5-loading-control-ab8245'>ab8245</a>) 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 2:
Western blot - Mouse SIRPA knockout RAW 264.7 cell line (<a href='/en-us/products/cell-lines/mouse-sirpa-knockout-raw-2647-cell-line-ab281618'>ab281618</a>)
Lane 3:
THP-1 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 55 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Mouse SIRPA knockout RAW 264.7 cell lysate (AB282969)
35 bp deletion in exon 6
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
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
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB214274
Alexa Fluor® 647 Anti-SIRP alpha antibody [EPR16264]
RabMAb|Recombinant
![Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-SIRP alpha antibody [EPR16264] (AB214274)](https://content.abcam.com/products/images/alexa-fluor-647-sirp-alpha-antibody-epr16264-ab214274--immunocytochemistry-immunofluorescence-img41393.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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