Anti-Homeobox protein cut-like 1 antibody [EPR26508-67]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Homeobox protein cut-like 1 with ab307821 at 1/50 dilution (9.66 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing nuclear staining in HeLa cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• IP

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Immunoprecipitation - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Homeobox protein cut-like 1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab307821 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307821 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa whole cell lysate 10 ug Lane 2 : ab307821 IP in HeLa whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307821 in hela whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Homeobox protein cut-like 1 with ab307821 at 1/1000 dilution (0.483 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab307821 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Homeobox protein cut-like 1 with ab307821 at 1/1000 dilution (0.483 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on mouse cerebrum (PMID : 20534841). The section was incubated with ab307821 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Homeobox protein cut-like 1 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab307821 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307821 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Neuro-2a whole cell lysate 10 ug Lane 2 : ab307821 IP in Neuro-2a whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307821 in Neuro-2a whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds.

• WB

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Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The Lanes 1 and 3 were developed using a high sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34997000). Lysates were freshly made and used immediately to minimize protein degradation. Exposure time : Lanes 1 and 3 : 180 seconds, lane 2 : 70 seconds.

All lanes:

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (ab307821) at 1/1000 dilution

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 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 90-240 kDa

false

• WB

Supplier Data

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34997000).

All lanes:

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (ab307821) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 10 µg

Lane 2:

HeLa transfected with siRNA specifically targeting CUTL1 whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 90-240 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

Exposure time : Lane 1 : 10 seconds; Lane 2 : 20 seconds; Lane 3 : 7 seconds.

All lanes:

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (ab307821) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

Rat cerebellum 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

Observed band size: 90-240 kDa

false

• WB

Supplier Data

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (AB307821)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34997000).

All lanes:

Western blot - Anti-Homeobox protein cut-like 1 antibody [EPR26508-67] (ab307821) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain 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 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 90-240 kDa

true

Exposure time: 180s