Anti-HCE antibody [EPR19384] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

HCE was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab201046 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab201046 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : SH-SY5Y whole cell lysate 10μg (Input).
Lane 2 : ab201046 IP in SH-SY5Y whole cell lysate.
Lane 3 : Rabbit IgG,monoclonal [EPR19384] -Isotype Control (ab172730) instead of ab201046 in SH-SY5Y whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-HCE antibody [EPR19384] (<a href='/en-us/products/primary-antibodies/hce-antibody-epr19384-ab201046'>ab201046</a>)

Predicted band size: 69 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on neurons of mouse cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HCE antibody [EPR19384] (<a href='/en-us/products/primary-antibodies/hce-antibody-epr19384-ab201046'>ab201046</a>) at 1/2000 dilution

Lane 1:

Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell 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: 69 kDa

Observed band size: 69 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HCE antibody [EPR19384] (<a href='/en-us/products/primary-antibodies/hce-antibody-epr19384-ab201046'>ab201046</a>) at 1/2000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 69 kDa

Observed band size: 69 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HCE antibody [EPR19384] (<a href='/en-us/products/primary-antibodies/hce-antibody-epr19384-ab201046'>ab201046</a>) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat kidney 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: 69 kDa

Observed band size: 69 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HCE antibody [EPR19384] - BSA and Azide free (AB251333)

This data was developed using ab201046, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HCE antibody [EPR19384] (<a href='/en-us/products/primary-antibodies/hce-antibody-epr19384-ab201046'>ab201046</a>) at 1/2000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast cell line) 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: 69 kDa

Observed band size: 69 kDa

false

Exposure time: 30s