Anti-Cystatin C antibody [EPR4413]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (AB109508)

Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab109508 at adilution of 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (AB109508)

Immunohistochemistry analysis of paraffin-embedded human kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on human kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (AB109508)

Immunohistochemistry analysis of paraffin-embedded rat kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on rat kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (AB109508)

Immunohistochemistry analysis of paraffin-embedded mouse kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on mouse kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Unknown

Western blot - Anti-Cystatin C antibody [EPR4413] (AB109508)

All lanes:

Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution

Lane 1:

Human testis lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

U87-MG cell lysate at 10 µg

Predicted band size: 15 kDa

Observed band size: 13 kDa

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• WB

Lab

Western blot - Anti-Cystatin C antibody [EPR4413] (AB109508)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : CST3 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : Caco2 whole cell lysate (20 μg)
Lane 4 : U20S whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109508 observed at 15 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109508 was shown to specifically react with CST3 when CST3 knockout samples were used. Wild-type and CST3 knockout samples were subjected to SDS-PAGE. ab109508 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 10000 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-Cystatin C antibody [EPR4413] (ab109508)

Predicted band size: 15 kDa,19 kDa

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• WB

Lab

Western blot - Anti-Cystatin C antibody [EPR4413] (AB109508)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST

Lane 1:

Western blot - Anti-Cystatin C antibody [EPR4413] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cystatin-c-antibody-epr4413-bsa-and-azide-free-ab217569'>ab217569</a>) at 1/10000 dilution

Lane 1:

Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution

All lanes:

Mouse thymus tissue lysate at 20 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: 15 kDa

Observed band size: 13 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-Cystatin C antibody [EPR4413] (AB109508)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST

Lane 1:

Western blot - Anti-Cystatin C antibody [EPR4413] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cystatin-c-antibody-epr4413-bsa-and-azide-free-ab217569'>ab217569</a>) at 1/10000 dilution

Lane 1:

Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution

All lanes:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 15 kDa

Observed band size: 13 kDa

false

Exposure time: 80s