Thermo Fisher Scientific p27kip1 Monoclonal Antibody (SX53G8)

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:200

-

Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

SX53G8

Immunogen

Purified glutathione S-transferase (GST)-p27Kip1 fusion protein

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

100 µg/mL

Purification

Protein A

Storage buffer

tris with NP-40, BSA

Contains

<0.1% sodium azide

Storage conditions

4° C

Shipping conditions

Wet ice

Product Specific Information

A recommended positive control tissue for this product is B-cell lymphoma, however positive controls are not limited to this tissue type.

The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.

p27Kip1, a candidate tumor suppressor gene, functions as a negative regulator of G1 progression and has been proposed to function as a possible mediator of TGF-β induced G1 arrest. Clone SX53G8 co-precipitates cdk4 in complex with p27Kip1 and is excellent for staining of formalin fixed tissues.

Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.

A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.

Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.

Target Information

p27 Kip1 functions as a negative regulator of G1 mouse expression and has been proposed to function as a possible mediator of TGFb induced G1 arrest. p27 Kip1 is an important regulator of cell cycle progression, a potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. p27 Kip1 forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. p27 Kip1 acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry. p27 Kip1 has a molecular weight of approximately 27kDa and the degradation of this protein, which is triggered by its CDK dependent phosphorylation and subsequent ubiquitination by SCF complexes, is required for the cellular transition from quiescence to the proliferative state.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.