Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis (PMID : 25145264; PMID : 20736371) is observed. The section was incubated with ab223256 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

Secondary antibody only control : Used PBS insread of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human stomach (PMID : 25145264) is observed. The section was incubated with ab223256 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• IP

Lab

Immunoprecipitation - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

Splicing factor 1 was immunoprecipitated from HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab223256 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed from the immunoprecipitate using ab223256 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab223256 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab223256 in HeLa whole cell lysate

Blocking/Dilution buffer : 5% NFDM/TBST.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-splicing factor 1 antibody [EPR22437-50] (<a href='/en-us/products/primary-antibodies/splicing-factor-1-antibody-epr22437-50-ab223256'>ab223256</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/splicing-factor-1-antibody-epr22437-50-ab223256'>ab223256</a> IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 68 kDa

Observed band size: 70 kDa,80 kDa

false

Exposure time: 15s

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis (PMID : 25145264) is observed. The section was incubated with ab223256 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in Neuro-2a cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Steroidogenic Factor 1/SF-1 with ab223256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat testis (PMID : 25145264) is observed. The section was incubated with ab223256 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223256).

• WB

Lab

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254199).

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 8752089; PMID : 17383426; PMID : 10103072; PMID : 25145264).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (<a href='/en-us/products/primary-antibodies/splicing-factor-1-antibody-epr22437-50-ab223256'>ab223256</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) 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: 68 kDa

Observed band size: 70 kDa,80 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

This data was developed using ab223256, the same antibody clone in a different buffer formulation.

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

This protein is easy to be degraded, to minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting right away.

All lanes:

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (<a href='/en-us/products/primary-antibodies/splicing-factor-1-antibody-epr22437-50-ab223256'>ab223256</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) frozen lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) fresh 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: 70-80 kDa

false

Exposure time: 7s

• WB

Lab

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] - BSA and Azide free (AB254199)

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 8752089; PMID : 17383426; PMID : 10103072; PMID : 25145264).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254199).

All lanes:

Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (<a href='/en-us/products/primary-antibodies/splicing-factor-1-antibody-epr22437-50-ab223256'>ab223256</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse testis 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

Observed band size: 70 kDa,80 kDa

false

Exposure time: 26s