AB275505

Human ANPEP (CD13) knockout THP-1 cell lysate

Name: Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505) | Abcam
Brand: Abcam Limited
SKU: AB275505
Price: 114000 JPY
Availability: InStock

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ANPEP KO cell lysate available now. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2.

別名を表示する

AMPN_HUMAN, ANPEP, AP-M, AP-N, Alanyl (membrane) aminopeptidase, Alanyl aminopeptidase, Aminopeptidase M, Aminopeptidase N, CD13 antigen, LAP 1, Microsomal aminopeptidase, Myeloid plasma membrane glycoprotein CD13, P150, PEPN, gp150, hAPN

4 Images

Lab

Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (AB275505)

Lane 1 : Wild-type THP-1 cell lysate 30 ug
Lane 2 : ANPEP knockout THP-1 cell lysate 30 ug
Lane 3 : PANC-1 cell lysate 30 ug
Lane 4 : HEK-293 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab108310 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108310 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and CD13 knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108310 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (<a href='/products/primary-antibodies/cd13-antibody-epr4058-ab108310'>ab108310</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 30 µg

Lane 2:

ANPEP knockout THP-1 cell lysate at 30 µg

Lane 2:

Western blot - Human ANPEP (CD13) knockout THP-1 cell line (<a href='/products/cell-lines/human-anpep-cd13-knockout-thp-1-cell-line-ab273759'>ab273759</a>)

Lane 3:

PANC-1 cell lysate at 30 µg

Lane 4:

HEK-293 cell lysate at 30 µg

Predicted band size: 109 kDa

Observed band size: 160 kDa

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Lab

Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (AB275505)

Lane 1 : Wild-type THP-1 cell lysate 30 ug
Lane 2 : ANPEP knockout THP-1 cell lysate 30 ug
Lane 3 : PANC-1 cell lysate 30 ug
Lane 4 : HEK-293 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab108382 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108382 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and CD13 knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab108382 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD13 antibody [EPR4059] (<a href='/products/primary-antibodies/cd13-antibody-epr4059-ab108382'>ab108382</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 30 µg

Lane 2:

ANPEP knockout THP-1 cell lysate at 30 µg

Lane 2:

Western blot - Human ANPEP (CD13) knockout THP-1 cell line (<a href='/products/cell-lines/human-anpep-cd13-knockout-thp-1-cell-line-ab273759'>ab273759</a>)

Lane 3:

PANC-1 cell lysate at 30 µg

Lane 4:

HEK-293 cell lysate at 30 µg

Predicted band size: 109 kDa

Observed band size: 160 kDa

false

Sanger Sequencing - Human ANPEP (CD13) knockout THP-1 cell lysate (AB275505)

Sanger seq

Supplier Data

Sanger Sequencing - Human ANPEP (CD13) knockout THP-1 cell lysate (AB275505)

Homozygous : 2 bp deletion in exon 2

Lab

Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (AB275505)

Lanes 1 - 4 : Merged signal (red and green). Green - ab227663 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab227663 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and ANPEP knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab227663 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD13 antibody [SP187] (<a href='/products/primary-antibodies/cd13-antibody-sp187-ab227663'>ab227663</a>) at 1/400 dilution

Lane 1:

Wild-type THP-1 cell lysate at 30 µg

Lane 2:

Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505) at 30 µg

Lane 3:

PANC-1 cell lysate at 30 µg

Lane 4:

HEK-293 cell lysate at 30 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 109 kDa

Observed band size: 160 kDa,37 kDa

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Key facts

細胞タイプ

THP-1

生物種

Human

組織

Blood

ノックアウト検証方法

Immunocytochemistry,Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2

疾病

Acute Monocytic Leukemia

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Reactivity data

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製品の詳細

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.

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製品内容

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出荷温度及び保存条件

遺伝子名

ANPEP

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Immunocytochemistry, Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

CD13 also known as aminopeptidase N (ANPEP) is a transmembrane protein with a molecular weight of approximately 150 kDa. It functions as a zinc-dependent metalloenzyme which cleaves N-terminal amino acids from peptides and proteins. CD13 protein is expressed on various cell types including myeloid cells epithelial cells endothelial cells and fibroblasts. Its presence is significantly observed in the brush border of the small intestinal mucosa and the renal proximal tubule. Researchers can study CD13 using anti-CD13 antibodies and CD13 ELISA kits.

Biological function summary

CD13 regulates peptide-mediated signaling and controls the maturation and catabolism of bioactive peptides. The enzymatic function of CD13 influences processes such as cell proliferation motility and angiogenesis. It does not operate as part of a larger complex but its activity modulates several cellular and systemic functions. CD13's role in these biological processes highlights its importance in modulating local and systemic peptide pools which contributes to its diverse physiological effects.

Pathways

CD13 plays significant roles in the renin-angiotensin system and the regulation of inflammatory responses. In the renin-angiotensin system CD13 modulates the activity of angiotensin influencing blood pressure and fluid balance. Through its enzymatic activity CD13 interacts with proteins such as ACE another critical player in this pathway. In inflammation CD13 regulates the availability of chemotactic peptides affecting leukocyte migration and adhesion. The proteins it works with in inflammatory pathways include cytokines which CD13 indirectly modulates by altering chemokine activity.

Aberrant CD13 expression and activity are implicated in cancer and inflammatory diseases. In cancer overexpression of CD13 correlates with tumor growth and metastasis particularly in cancers like renal cell carcinoma and prostate cancer. It can confer an increased malignant phenotype by promoting angiogenesis and immune evasion. In inflammatory disorders dysregulation of CD13 exacerbates diseases such as rheumatoid arthritis through its impact on peptide-mediated signaling and immune cell movement. Its connection with inflammatory cytokines like IL-8 indicates its important role in mediating inflammatory responses.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human ANPEP (CD13) knockout THP-1 cell lysate