Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)

Rabbit Monoclonal CSK antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), WB, IHC-P, IP and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (AB300133), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	Flow Cyt (Intra)	WB	IHC-P	IP
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Tested	Tested	Tested
Rat	Not recommended	Not recommended	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information CSK

Function

Non-receptor tyrosine-protein kinase that plays an important role in the regulation of cell growth, differentiation, migration and immune response. Phosphorylates tyrosine residues located in the C-terminal tails of Src-family kinases (SFKs) including LCK, SRC, HCK, FYN, LYN, CSK or YES1. Upon tail phosphorylation, Src-family members engage in intramolecular interactions between the phosphotyrosine tail and the SH2 domain that result in an inactive conformation. To inhibit SFKs, CSK is recruited to the plasma membrane via binding to transmembrane proteins or adapter proteins located near the plasma membrane. Suppresses signaling by various surface receptors, including T-cell receptor (TCR) and B-cell receptor (BCR) by phosphorylating and maintaining inactive several positive effectors such as FYN or LCK.

Alternative names

Tyrosine-protein kinase CSK, C-Src kinase, Protein-tyrosine kinase CYL, CSK

Recommended products

Rabbit Monoclonal CSK antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), WB, IHC-P, IP and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR24673-97
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Aliquoting information: Upon delivery aliquot

Notes

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Western blot - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
This data was developed using the same antibody clone in a different buffer formulation (Anti-CSK antibody [EPR24673-97] ab300132 ).
Western blot: Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-CSK antibody [EPR24673-97] ab300132 was shown to bind specifically to CSK. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in CSK knockout cell line. To generate this image, wild-type and CSK knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: CSK knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human CSK knockout A549 cell line (Human CSK knockout A549 cell line ab300902)
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/5000 (0.111 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control: no staining on human cardiac muscle. The section was incubated with Anti-CSK antibody [EPR24673-97] ab300132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Western blot - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS was used as a blocking and diluting buffer. Lanes 1-4: Merged signal (red and green). Green - Anti-CSK antibody [EPR24673-97] ab300132 observed at 50 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa. Anti-CSK antibody [EPR24673-97] ab300132 Anti-CSK antibody [EPR24673-97] was shown to specifically react with CSK in wild-type HAP1 cells. Loss of signal was observed when knockout cell line was used. Wild-type and CSK knockout samples were subjected to SDS-PAGE. Anti-CSK antibody [EPR24673-97] ab300132 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: CSK knockout HAP1 whole cell lysate at 20 µg
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg
Secondary
All lanes: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS at 1/10000 dilution
Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
CSK was immunoprecipitated from 0.35 mg rat lymph node tissue lysate 10 µg with Anti-CSK antibody [EPR24673-97] ab300132 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CSK antibody [EPR24673-97] ab300132 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1 (input): Rat lymph node tissue lysate 10 µg. Lane 2 (+): Anti-CSK antibody [EPR24673-97] ab300132 in Rat lymph node tissue lysate. Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in rat lymph node tissue lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 3 seconds. The identity of the lower MW band at approximately 37kDa is unknown.
All lanes: Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/30 dilution
Lane 1: Rat lymph node tissue lysate at 10 µg
Lane 2: Anti-CSK antibody [EPR24673-97] ab300132 in Rat lymph node tissue lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in rat lymph node tissue lysate
Observed band size: 50 kDa
Flow Cytometry (Intracellular) - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell, Right) / CSK knockout HAP1(Left) cells labelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/5000 (0.111 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat spleen. The section was incubated with Anti-CSK antibody [EPR24673-97] ab300132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: PBS was used instead of primary antibody (Anti-CSK antibody [EPR24673-97] ab300132) followed by ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/5000 (0.111 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen. The section was incubated with Anti-CSK antibody [EPR24673-97] ab300132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: PBS was used instead of primary antibody (Anti-CSK antibody [EPR24673-97] ab300132) followed by ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/5000 (0.111 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human tonsil (PMID: 9287362). The section was incubated with Anti-CSK antibody [EPR24673-97] ab300132 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Western blot - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/1000 dilution
Lane 1: Human tonsil tissue lysate at 20 µg
Lane 2: Mouse spleen tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Exposure time: 3s
Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
CSK was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma b lymphocyte) whole cell lysate 10 µg with Anti-CSK antibody [EPR24673-97] ab300132 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CSK antibody [EPR24673-97] ab300132 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1 (Input): Ramos (human burkitt's lymphoma b lymphocyte) whole cell lysate 10 µg. Lane 2 (+): Anti-CSK antibody [EPR24673-97] ab300132 in Ramos whole cell lysate. Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in Ramos whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 3 seconds
All lanes: Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/30 dilution
Lane 1: Ramos (human burkitt's lymphoma b lymphocyte) whole cell lysate at 10 µg
Lane 2: Anti-CSK antibody [EPR24673-97] ab300132 in Ramos whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in Ramos whole cell lysate
Observed band size: 50 kDa
Immunocytochemistry - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Immunofluorescent analysis of 80% methanol-fixed, 0.1% TritonX-100 permeilized Jurkat (human T cell leukemia T lymphocyte) cells lebelling CSK with Anti-CSK antibody [EPR24673-97] ab300132 at 1/50 (11.12 µg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Jurkat cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
Western blot - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: Lanes 1-3: 15 seconds; Lanes 4-7: 3 minutes; Lanes 8-9: 15 seconds.
All lanes: Western blot - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/1000 dilution
Lane 1: Ramos (Human burkitt's lymphoma b lymphocyte) whole cell lysate at 1/20 dilution
Lane 2: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 1/20 dilution
Lane 3: RAW 264.7 (Mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 1/20 dilution
Lane 4: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 1/20 dilution
Lane 5: C6 (Rat glial tumor glial cell) whole cell lysate at 1/20 dilution
Lanes 6 - 7: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 1/20 dilution
Lane 8: Mouse lymph node tissue lysate at 1/20 dilution
Lane 9: Rat lymph node tissue lysate at 1/20 dilution
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (BSA and Azide free) (ab300133)
CSK was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 µg with Anti-CSK antibody [EPR24673-97] ab300132 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CSK antibody [EPR24673-97] ab300132 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1 (input): RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 µg. Lane 2 (+): Anti-CSK antibody [EPR24673-97] ab300132 in RAW 264.7 whole cell lysate. Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in RAW 264.7 whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 3 seconds
All lanes: Immunoprecipitation - Anti-CSK antibody [EPR24673-97] (Anti-CSK antibody [EPR24673-97] ab300132) at 1/30 dilution
Lane 1: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg
Lane 2: Anti-CSK antibody [EPR24673-97] ab300132 in RAW 264.7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CSK antibody [EPR24673-97] ab300132 in RAW 264.7 whole cell lysate
Observed band size: 50 kDa