SP1 Monoclonal antibody

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  WB analysis using 66508-1-Ig

  Various cells were subjected to SDS PAGE followed by western blot with 66508-1-Ig (SP1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of NIH/3T3 using 66508-1-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 66508-1-Ig (SP1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IHC staining of human lung cancer using 66508-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66508-1-Ig (SP1 antibody) at dilution of 1:350 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 66508-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66508-1-Ig (SP1 antibody) at dilution of 1:350 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 66508-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66508-1-Ig (SP1 antibody) at dilution of 1:350 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 66508-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66508-1-Ig (SP1 antibody) at dilution of 1:350 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of human colon cancer using 66508-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human colon cancer tissue using SP1 antibody (66508-1-Ig, Clone: 4G3H3 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of HEK-293 using 66508-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using SP1 antibody (66508-1-Ig, Clone: 4G3H3 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of HEK-293 using 66508-1-Ig

  1X10^6 HEK-293 cells were intracellularly stained with 0.4 ug Anti-Human SP1 (66508-1-Ig, Clone:4G3H3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

SP1 Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA

Cat No. 66508-1-Ig

Clone No.4G3H3

Publications(6)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA

Synonyms

4G3H3, SP 1, Sp1 transcription factor, Specificity protein 1, TSFP1

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Tested Applications

• Positive WB detected in: Jurkat cells, NIH/3T3 cells, HEK-293 cells, HeLa cells
• Positive IHC detected in: human lung cancer tissue, human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human colon cancer tissue
• Positive IF/ICC detected in: HEK-293 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunohistochemistry (IHC): IHC : 1:175-1:700
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 4 publications below
• IHC: See 1 publications below
• IF: See 1 publications below

Product Information

66508-1-Ig targets SP1 in WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: SP1 fusion protein Ag16720
• Full Name: Sp1 transcription factor
• Calculated Molecular Weight: 785 aa, 81 kDa
• Observed Molecular Weight: 95-105 kDa, 60-65 kDa
• GenBank Accession Number: BC062539
• Gene Symbol: SP1
• Gene ID (NCBI): 6667
• ENSEMBL Gene ID: ENSG00000185591
• RRID: AB_2881871
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: P08047
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The transcription factor Sp1 is a C2H2 zinc-finger protein that is involved in the regulation of a wide variety of genes, including housekeeping genes and tumor-developing genes. It is associated with tumor development, growth, and metastasis [PMID: 16209919]. It regulates the expression of a large number of genes involved in a variety of processes such as cell growth, apoptosis, differentiation and immune responses. Besides, it has a role in modulating the cellular response to DNA damage, recruiting SMARCA4/BRG1 on the c-FOS promoter, regulation of FE65 gene expression [PMID:11371615,16332679]. SP1 has some isoforms with MW ~95-106 kDa and ~60 kDa [PMID:17462816].

Publications

Species	Application	Title
rat	WB	Br J Pharmacol; Sarco/endoplasmic reticulum Ca 2+ -ATPase 2b mediates oxidation-induced endoplasmic reticulum stress to regulate neuropathic pain; Authors - Shaoheng Li
mouse	IHC	Evid Based Complement Alternat Med; Guifu Dihuang Pills Ameliorated Mucus Hypersecretion by Suppressing Muc5ac Expression and Inactivating the ERK-SP1 Pathway in Lipopolysaccharide/Cigarette Smoke-Induced Mice; Authors - Huanhuan Zhang
human	WB	Endocrinology; AURKA enhances the glycolysis and development of ovarian endometriosis through ERβ; Authors - Yujun Sun
human	WB	Acta Biochim Biophys Sin (Shanghai); Propofol protects against high glucose-mediated endothelial injury via inhibition of COX2 and iNOS expressions; Authors - Jiayun Shao
human	WB	Genomics; STRIP2 is regulated by the transcription factor Sp1 and promotes lung adenocarcinoma progression via activating the PI3K/AKT/mTOR/MYC signaling pathway; Authors - Junfan Pan
	IF	EMBO Rep; Histone modifications and Sp1 promote GPR160 expression in bone cancer pain within rodent models; Authors - Chengfei Xu