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LIF Antibody (Center) 精选

Affinity Purified Rabbit Polyclonal Antibody (Pab)

Immunohistochemical analysis of paraffin-embedded human kidney tissue using AP6981C performed on the Leica® BOND RXm. Samples were incubated with primary antibody(1/500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human kidney tissue using AP6981C performed on the Leica® BOND RXm. Samples were incubated with primary antibody(1/500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human brain tissue using AP6981C performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature; antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody(1:1000) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
Overlay histogram showing A431 cells stained with AP6981C(green line). The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at Room temperature. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Overlay histogram showing U-2 OS cells stained with AP6981C(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP6981C, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(1583138) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Overlay histogram showing U-2 OS cells stained with AP6981C(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP6981C, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(1583138) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Anti-LIF Antibody (Center) at 1:2000 dilution + Human spleen lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 22 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-LIF Antibody (Center) at 1:1000 dilution + Rat spleen lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 25 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes : Anti- LIF Antibody (Center) at 1:1000 dilution Lane 1: KG-1 whole cell lysate Lane 2:Human spleen whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 22 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-LIF Antibody (Center) at 1:2000 dilution + KG-1 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 22 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot analysis of lysates from rat PC-12 cell line, mouse spleen tissue lysate (from left to right), using LIF Antibody (Center)(Cat. #AP6981c). AP6981c was diluted at 1:1000 at each lane. A goat anti-rabbit IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysates at 20ug per lane.
Western blot analysis of LIF Antibody (Center) (Cat. #AP6981c) in CEM cell line lysates (35ug/lane). LIF (arrow) was detected using the purified Pab.
Western blot analysis of LIF (arrow) using rabbit polyclonal LIF Antibody (Center) (Cat. #AP6981c). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected (Lane 2) with the LIF gene.
Formalin-fixed and paraffin-embedded human colon carcinoma reacted with LIF Antibody (Center), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Flow cytometric analysis of CEM cells using LIF Antibody (Center)(bottom histogram) compared to a negative control cell (top histogram). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, FC, IHC-P-Leica, E
Primary Accession	P15018
Reactivity	Human, Rat, Mouse
Predicted	Bovine, Canine, Rabbit
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	22008 Da
Antigen Region	72-101 aa

Additional Information
Gene ID	3976
Other Names	Leukemia inhibitory factor, LIF, Differentiation-stimulating factor, D factor, Melanoma-derived LPL inhibitor, MLPLI, Emfilermin, LIF, HILDA
Target/Specificity	This LIF antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 72-101 amino acids from the Central region of human LIF.
Dilution	WB~~1:1000 FC~~1:10~50 IHC-P-Leica~~1:1000 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	LIF Antibody (Center) is for research use only and not for use in diagnostic or therapeutic procedures.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	LIF (HGNC:6596)
Synonyms	HILDA
Function	Functions as a cytokine that binds to the LIF receptor complex (PubMed:11294841, PubMed:14527405, PubMed:36930708). The LIF receptor complex indeed consists of two signaling receptor subunits IL6ST/gp130 and LIFR that transduce the signal into the cell (PubMed:14527405, PubMed:36930708). Functionally, regulates cell self- renewal, differentiation and survival of embryonic stem cells (ESCs) and is crucial for embryonic implantation (By similarity). Mechanistically, ligand binding to LIFR, induces heterodimerization with IL6ST/gp130 activating JAK tyrosine kinases (JAK1 or JAK2 and to a lesser extent TYK2) bound to their intracellular domains (PubMed:11294841). These kinases subsequently phosphorylate IL6ST/gp130 and LIFR (PubMed:11294841). The tyrosine phosphorylated signaling receptors serve in turn as docking sites for recruitment and activation of signal transducers and activators of transcription (STAT3 and to lesser extent STAT1) (PubMed:11294841). In parallel to the activation of the STAT3-pathway, the binding of LIF to the LIF receptor complex leads to the activation of the mitogen-activated protein kinase (MAPK) and the phosphatidylinositol 3-kinase (PI(3)K) pathways (PubMed:11294841). Tyrosine phosphorylation of the p85 subunit of PI(3)K by JAK1 leads to activation of the serine/ threonine kinase AKT (protein kinase B) (PubMed:11294841). PI(3)K-dependent signaling is required for efficient self-renewal of murine ES cells, and regulation of ERK activity is functionally important in this response (By similarity). Moreover, induces tyrosine phosphorylation of PTPN11, that associates with IL6ST/gp130-LIFR, which in turn recruits the p85 regulatory subunit of PI(3)K protein kinase, promoting PI3K-AKT activation, and GRB2-RAS-MAPK signaling (PubMed:11294841).
Cellular Location	Secreted.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

LIF is a pleiotropic cytokine with roles in several different systems. It is involved in the induction of hematopoietic differentiation in normal and myeloid leukemia cells, induction of neuronal cell differentiation, regulator of mesenchymal to epithelial conversion during kidney development, and may also have a role in immune tolerance at the maternal-fetal interface.

REFERENCES

Novotny,Z., et.al., Folia Biol. (Praha) 55 (3), 92-97 (2009)

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¥ 1,250.00

Cat# AP6981C

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LIF Antibody (Center) 精选

Affinity Purified Rabbit Polyclonal Antibody (Pab)

BACKGROUND

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