Anti-CD13 antibody [EPR4058]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human prostatic carcinoma tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling CD13 with purified ab108310 at 1/1600 dilution (0.43 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human normal liver tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human lung adenocarcinoma tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human normal tonsil tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human normal colon tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [EPR4058] (AB108310)

Confocal image showing membranous staining in THP-1 cells

ab108310 (purified) at 1/100 staining CD13 in the THP-1 (human monocytic leukemia monocyte) cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol. Samples were incubated with primary antibody 1/500. ab150077 An Alexa Fluor® 488-conjugated Goat anti-Rabbit IgG at 1/1000 was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 was used as a counter stain and DAPI was used as a nuclear counter stain.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 staining CD13 in wild-type THP-1 cells (top panel) and ANPEP knockout THP-1 cells (bottom panel) (ab273759). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108310 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) at 1/250 staining CD13 in human kidney tissue by immunohistochemistry. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human astrocytoma tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human hepatocellular carcinoma tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human normal stomach tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

ab108310 (unpurified) showing positive staining in human normal breast tissue. Heat mediated antigen retrieval was performed using citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD13 antibody [EPR4058] (AB108310)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling CD13 with purified ab108310 at 1/1600 dilution (0.43 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

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 ANPEP 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] (ab108310) 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='/en-us/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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• WB

Lab

Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : Empty
Lane 3 : CD13 knockout HAP1 whole cell lysate (20 μg)
Lane 4 : Empty
Lane 5 : HeLa whole cell lysate (20 μg)

Lanes 1 - 5 : Merged signal (red and green). Green - ab108310 (unpurified) observed at 160 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab108310 was shown to recognize CD13 when CD13 knockout samples were used, along with additional cross-reactive bands. Wild-type and CD13 knockout samples were subjected to SDS-PAGE. ab108310 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (ab108310)

Predicted band size: 109 kDa

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• WB

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Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (ab108310) at 1/1000 dilution

Lane 1:

THP-1 cell lysate at 10 µg

Lane 2:

human fetal liver lysate

Predicted band size: 109 kDa

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• WB

Lab

Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (ab108310) at 1/5000 dilution

Lane 1:

THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg

Lane 2:

Rat kidney lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 109 kDa

Observed band size: 150 kDa

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• WB

Lab

Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the lower MW band at approximately 75 kDa is non-specific banding.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Anpep/CD13-KO homozygous mice (Strain ID : T027669).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (ab108310) at 1/1000 dilution

Lane 1:

Wild-type mouse liver tissue lysate (male)

Lanes 2 - 3:

Anpep/CD13 knockout liver tissue lysate (male)

Lane 4:

Wild-type mouse kidney tissue lysate (male)

Lanes 5 - 6:

Anpep/CD13 knockout kidney tissue lysate (male)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 150 kDa

false

Exposure time: 38s

• WB

Lab

Western blot - Anti-CD13 antibody [EPR4058] (AB108310)

All lanes:

Western blot - Anti-CD13 antibody [EPR4058] (ab108310) at 1/20000 dilution

All lanes:

Mouse kidney lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 109 kDa

Observed band size: 150 kDa

false