REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H | Endogenous | Rabbit IgG |
Immunofluorescent analysis of Ramos cells untreated (left) or treated with IgM (12 μg/mL, 10 min, right) using Phospho-HS1 (Tyr397) (D12C1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (red). Blue pseudocolor= ProLong® Gold Antifade Reagent with DAPI #8961.
Learn more about how we get our imagesFlow cytometric analysis of Ramos cells, untreated (blue) or treated with IgM (12 μg/ml, 10 min; green), using Phospho-HS1 (Tyr397) (D12C1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (dashed lines).
Learn more about how we get our imagesNOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
posted July 2009
revised June 2017
Protocol Id: 407
Application | Dilutions |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:100 |
Flow Cytometry | 1:50 |
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Phospho-HS1 (Tyr397) (D12C1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of HS1 protein only when phosphorylated at Tyr397.
Human
Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr405 of mouse HS1 protein. This site corresponds to Tyr397 of human HS1 protein.
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-HS1 (Tyr397) (D12C1) XP® Rabbit mAb #8714.
HS1 (HCLS1, LckBP1, p75) is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (1,2). HS1 contains four cortactin repeats and a single SH3 domain (2). This intracellular protein is phosphorylated following immune receptor activation, which promotes recruitment of HS1 to the immune synapse (3-5). Phosphorylation of HS1 is required to regulate actin dynamics and provide docking sites for many other signaling molecules, such as Vav1 and PLCγ1 (6). HS1 also plays an important role in platelet activation (7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Alexa Fluor is a registered trademark of Life Technologies Corporation. The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
Explore pathways related to this product.
Product # | Size | Price |
---|---|---|
13708S | 100 µl (50 tests) | $ 356.0 |