AM26430AF-N Ly6A/E / SCA1 antibody

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0.1 mg / €390.00
Please visit the country specific website of OriGene Technologies or contact your local Distributor to buy this product.

Quick Overview

Rat anti Mouse Ly6A/E / SCA1 238B

Product Description for Ly6A/E / SCA1

Rat anti Mouse Ly6A/E / SCA1 238B.
Presentation: Azide Free
Product is tested for Flow Cytometry.

Properties for Ly6A/E / SCA1

Product Category Primary Antibodies
Target Category
Quantity 0.1 mg
Synonyms Ly-6A.2/Ly-6E.1, Lymphocyte antigen 6A-2/6E-1, SCA-1, Stem cell antigen 1, T-cell-activating protein, TAP
Presentation Azide Free
Reactivity Ms
Applications F
Clonality Monoclonal
Clone 238B
Host Rat
Isotype IgG2a
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer OriGene Technologies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Swiss Prot Num:
P05533
Immunogen:
Mouse Sca-1 transfected LO cells
GeneID:
110454
Isotype control SM26A
Add. information This product was originally produced by MBL International.
Application Flow cytometry: 10 μg/ml. For details see protocol below.
Background Sca-1 is a member of the Ly-6 antigen family which molecular mass of 8 kDa under nonreducing conditions and of 18 kDa under reducing conditions. Mouse hematopoietic stem cell expresses low levels of Thy-1 antigen (Thy-1lo) and to be lineage-negative (Lin-); not express markers characteristic of B cells (B220), granulocytes (Gr-1), myelomonocytic cells (Mac-1) and T lymphocytes (CD4 and CD8). Recently, new monoclonal antibody, anti-Sca-1, was used to purify stem cells from the Thy-1lo, Lin- bone marrow subpopulation. Thy-1lo, Lin-, Sca-1+ (but not the Thy-1lo, Lin-, Sca-1-) population of bone marrow cells are highly purified pluripotent stem cells. They read out with nearly unit efficiency in assays for primitive myeloerythroid and thymic progenitor, and have a capability to admit lethally irradiated mouse to survive and be restored in all blood-cell lineages. The Thy-1lo, Lin-, Sca-1+ subpopulation thought to have all stem cells present in the bone marrow.
Protocols

Flow cytometric analysis for adherent cells
We usually use Fisher tubes or equivalents as reaction tubes for all steps described below.
1) Detach the cells from culture dish by cell dissociation buffer.
2) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3].
3) Resuspend the cells with washing buffer (5x10e6 cells/mL).
4) Add 50 µL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
5) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0.1% NaN3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature.
6) Add 40 µL of the primary antibody at the concentration as suggest in the APPLICATIONS diluted in the washing buffer. Mix well and incubate for 30 minutes at room temperature.
7) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
8) Add 30 µL of 1:40 FITC conjugated anti-rat IgG diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature.
9) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
10) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer.
(Positive controls for Flow cytometry; LO, lymphocyte, splenocyte)

General Readings
  1. Petersen, B. E., et al., Hepatology 37, 632-640 (2003).
Storage

Store (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Format
Purification:
Protein G agarose
Buffer System:
PBS containing 50% glycerol, pH 7.2. No preservative is contained.
State:
Liquid Ig fraction
Azide Free
Species Reactivity
Species reactivity (tested):
Mouse
Specificity
Specificity:
This antibody reacts with Sca-1.
Gene ID 110454

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