Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22437-50] to splicing factor 1
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-splicing factor 1 antibody [EPR22437-50]
See all splicing factor 1 primary antibodies
Description

Rabbit monoclonal [EPR22437-50] to splicing factor 1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- ICC/IF: HeLa and Neuro-2a cells. WB: HeLa, RAW 264.7, NIH/3T3 and PC-12 whole cell lysate; mouse and rat testis lysate; human, mouse and rat brain cell lysate; rat heart cell lysate. IHC-P: Human testis and stomach tissue; mouse and rat testis tissue. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22437-50
Isotype

IgG
Research areas

Immunocytochemistry/ Immunofluorescence - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling splicing factor 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.
Immunocytochemistry/ Immunofluorescence - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling splicing factor 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling splicing factor 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.
Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

All lanes : Anti-splicing factor 1 antibody [EPR22437-50] (ab223256) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : Mouse testis cell lysate
Lane 3 : Rat testis cell lysate
Lane 4 : Human brain cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 52 kDa
Observed band size: 75 kDa
why is the actual band size different from the predicted?

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

The lower bands represent different Splicing factor 1 isoforms and/or degraded fragments.

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-3: 10 secs; Lane 4: 81 secs.
Flow Cytometry - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling splicing factor 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.
Immunoprecipitation - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Splicing factor 1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab223256 at 1/30 dilution. 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 whole cell lysate 10 µg (Input).
Lane 2: ab223256 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223256 in HeLa whole cell lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 1 second.

The lower bands below 75kDa represent Splicing factor 1 short isoforms and/or degraded fragments.
Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Anti-splicing factor 1 antibody [EPR22437-50] (ab223256) at 1/1000 dilution + Mouse brain lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 52 kDa

Exposure time: 15 seconds

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:8752089; PMID: 17383426; PMID: 10103072; PMID: 25145264). The lower active bands represent different Splicing factor 1 isoforms and/or degraded fragments.

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

All lanes : Anti-splicing factor 1 antibody [EPR22437-50] (ab223256) at 1/1000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Rat heart cell lysate
Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 5 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 52 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:8752089; PMID: 17383426; PMID:10103072; PMID: 25145264). The lower bands represent different Splicing factor 1 isoforms and/or degraded fragments.

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2: 10 secs; Lanes 3-5: 26 secs.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling splicing factor 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling splicing factor 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling splicing factor 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.
Anti-splicing factor 1 antibody [EPR22437-50] (ab223256)

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