重组Anti-hnRNP H抗体[EPR25302-12]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling hnRNP H with ab289974 at 1/500 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on human testis. The section was incubated with ab289974 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 LeicaDS9800 (Bond™, Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling hnRNP H with ab289974 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunohistochemical analysis of paraffin-embedded Human prostatic hype tissue labelling hnRNP H with ab289974 at 1/500 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on human prostatic hyperplasia. The section was incubated with ab289974 for 30 mins at room temperatureThe immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™, Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Intracellular flow analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling hnRNP H with ab289974 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Lab

Immunoprecipitation - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

hnRNP H was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate with ab289974 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289974 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate 10 μg

Lane 2 : ab289974 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289974 in HeLa whole cell lysate

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

Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-hnRNP H antibody [EPR25302-12] (ab289974)

Predicted band size: 49 kDa

Observed band size: 50 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling hnRNP H with ab289974 at 1/500 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab289974 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 LeicaDS9800 (Bond™, Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling hnRNP H with ab289974 at 1/500 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab289974 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 LeicaDS9800 (Bond™, Polymer Refine Detection) .

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling hnRNP H with ab289974 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells llabeling hnRNP H with ab289974 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in C2C12 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

hnRNP H was immunoprecipitated from 0.35 mg C2C12 (Mouse myoblasts myoblast), whole cell lysate 10 µg with ab289974 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289974 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : C2C12 (Mouse myoblasts myoblast), whole cell lysate 10 µg

Lane 2 : ab289974 IP in C2C12 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289974 in C2C12 whole cell lysate

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

Exposure time : 5.5 seconds

All lanes:

Immunoprecipitation - Anti-hnRNP H antibody [EPR25302-12] (ab289974)

Predicted band size: 49 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23667175, PMID : 21915099)

Exposure times : Lane 1 : 3 minutes Lane 2-6 : 15 seconds

All lanes:

Western blot - Anti-hnRNP H antibody [EPR25302-12] (ab289974) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Rat liver tissue lysate at 20 µg

Lane 6:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23667175, PMID : 21915099)

Exposure times : Lane 1-3 : 3 seconds Lane 4-6 : 5 seconds

All lanes:

Western blot - Anti-hnRNP H antibody [EPR25302-12] (ab289974) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2OS(human bone osteosarcoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

Lane 4:

C2C12 (Mouse myoblasts myoblast), whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23667175, PMID : 21915099)

Exposure time : 8 seconds

All lanes:

Western blot - Anti-hnRNP H antibody [EPR25302-12] (ab289974) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti hnRNP H whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-hnRNP H antibody [EPR25302-12] (AB289974)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody reacts with HNRPH1 and HNRPH2, but does not react with HNRNPF and HNRPH3. Exposure time : 114 seconds

All lanes:

Western blot - Anti-hnRNP H antibody [EPR25302-12] (ab289974) at 1/1000 dilution

Lane 1:

Myc-tagged human HNRPH1 (HNRNPH1) (NM_005520) recombinant protein (20 ng)

Lane 2:

Myc-tagged human HNRPH2 (HNRNPH2) (NM_019597) recombinant protein (20 ng)

Lane 3:

Myc-tagged human hnRNP F (HNRNPF) (NM_001098207) recombinant protein(20 ng)

Lane 4:

Myc-tagged human HNRPH3 (HNRNPH3) (NM_021644) recombinant protein (20 ng)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 50 kDa

false