Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

This data was developed using ab92741, the same antibody clone in a different buffer formulation.

ab92741 at 1/100 dilution staining p27 KIP in Mouse thymus by Immunohistochemistry, Paraffin-embedded tissue.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

This data was developed using ab92741, the same antibody clone in a different buffer formulation.

ab92741 at 1/100 dilution staining p27 KIP in Mouse lung by Immunohistochemistry, Paraffin-embedded tissue.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

This data was developed using ab92741, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling p27 KIP 1 with ab92741 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in Leydig and Sertoli cells and Leydig cells of rat testis is observed (PMID : 10098522). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody.

Heat mediated antigen retrieval was performed using Universal HIER antigen retrieval reagent (10X) (ab208572).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

This data was developed using ab92741, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p27 KIP 1 with ab92741 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in Leydig and Sertoli cells and Leydig cells of mouse testis is observed (PMID : 10098522). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody.

Heat mediated antigen retrieval was performed using Universal HIER antigen retrieval reagent (10X) (ab208572).

• WB

Unknown

Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

This data was developed using ab92741, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (<a href='/en-us/products/primary-antibodies/p27-kip-1-antibody-epfhcr16-ab92741'>ab92741</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 cell lysate at 10 µg

Lane 2:

C6 cell lysate at 10 µg

Lane 3:

Neuro-2a cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP-conjugated at 1/2000 dilution

Predicted band size: 22 kDa

Observed band size: 27 kDa

false

• WB

Lab

Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] - BSA and Azide free (AB247597)

False colour image of Western blot : Anti-p27 KIP 1 antibody [EPFHCR16] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab92741 was shown to bind specifically to p27 KIP 1. A band was observed at 28 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CDKN1B knockout cell line ab281619 (knockout cell lysate ab282970). To generate this image, wild-type and CDKN1B 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-p27 KIP 1 antibody [EPFHCR16] (<a href='/en-us/products/primary-antibodies/p27-kip-1-antibody-epfhcr16-ab92741'>ab92741</a>) at 1/1000 dilution

Lane 1:

Wild-type RAW 264.7 cell lysate at 20 µg

Lane 2:

CDKN1B knockout RAW 264.7 cell lysate at 20 µg

Lane 2:

Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (<a href='/en-us/products/cell-lines/mouse-cdkn1b-p27-kip-1-knockout-raw-2647-cell-line-ab281619'>ab281619</a>)

Predicted band size: 22 kDa

Observed band size: 28 kDa

false