Septin 2 Monoclonal antibody

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  WB analysis using 60075-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Alpha Tubulin Monoclonal antibody (HRP-66031) as loading control.

• 

  WB analysis of HEK-293 using 60075-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 Antibody) at various dilutions incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 60075-1-Ig

  WB result of Septin 2 antibody (60075-1-Ig; 1:30000; incubated at room temperature for 1.5 hours) with sh-Control and sh-Septin 2 transfected HEK-293 cells.

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  WB analysis of HeLa using 60075-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis of human brain using 60075-1-Ig

  human brain tissue were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human testis using 60075-1-Ig

  human testis tissue were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of fetal human brain using 60075-1-Ig

  fetal human brain tissue were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60075-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 60075-1-Ig

  Jurkat cells were subjected to SDS PAGE followed by western blot with 60075-1-Ig (Septin 2 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human renal cell carcinoma using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma tissue slide using 60075-1-Ig (Septin 2 Antibody) at dilution of 1:800 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human renal cell carcinoma using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma tissue slide using 60075-1-Ig (Septin 2 Antibody) at dilution of 1:800 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer using 60075-1-Ig(Septin 2 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human liver cancer using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer using 60075-1-Ig(Septin 2 antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human brain using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human brain using 60075-1-Ig(Septin 2 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human brain using 60075-1-Ig

  Immunohistochemical analysis of paraffin-embedded human brain using 60075-1-Ig(Septin 2 antibody) at dilution of 1:50 (under 40x lens).

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  IF Staining of hTERT-RPE cells using 60075-1-Ig

  IF result of anti-Septin 2 (60075-1-Ig, 1:1000) with hTERT-RPE cell by Dr. Moshe Kim. Green: Septin 2. Blue: DNA. 1% PFA fixed.

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  IF Staining of K-562 using 60075-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed K-562 cells using Septin 2 antibody (60075-1-Ig, Clone: 2F3B2 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of K-562 using 60075-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed K-562 cells using Septin 2 antibody (60075-1-Ig, Clone: 2F3B2 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1).

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  FC experiment of K-562 using 60075-1-Ig

  1X10^6 K-562 cells were intracellularly stained with 0.4 ug Anti-Human Septin 2 (60075-1-Ig, Clone:2F3B2) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

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  Affinity and Kinetic Characterization of 60075-1-Ig

  Biolayer interferometry (BLl) kinetic assays of 60075-1-Ig against Human SEPT2 were performed. The Affinity Constant is 1.23 nM.

• Featured Product
• KD/KO Validated

Septin 2 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 60075-1-Ig

Clone No.2F3B2

Publications(18)

Reviews (2)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA

Synonyms

DIFF6, hNedd5, KIAA0158, NEDD 5, NEDD5

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Tested Applications

• Positive WB detected in: LNCaP cells, Jurkat cells, fetal human brain tissue, human testis tissue, HeLa cells, HEK-293 cells, HepG2 cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, 4T1 cells, MCF-7 cells, HSCT6 cells, ROS1728 cells, Neuro-2A cells
• Positive IHC detected in: human renal cell carcinoma tissue, human brain tissue, human liver 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/ICC detected in: K-562 cells, hTERT-RPE cells
• Positive FC (Intra) detected in: K-562 cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:50-1:2000
• 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

• KD/KO: See 3 publications below
• WB: See 14 publications below
• IF: See 7 publications below
• CoIP: See 1 publications below

Product Information

60075-1-Ig targets Septin 2 in WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, canine
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag1946

  Product name: Recombinant human SEPT2 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 1-361 aa of BC014455

  Sequence: MSKQQPTQFINPETPGYVGFANLPNQVHRKSVKKGFEFTLMVVGESGLGKSTLINSLFLTDLYPERVISGAAEKIERTVQIEASTVEIEERGVKLRLTVVDTPGYGDAINCRDCFKTIISYIDEQFERYLHDESGLNRRHIIDNRVHCCFYFISPFGHGLKPLDVAFMKAIHNKVNIVPVIAKADTLTLKERERLKKRILDEIEEHNIKIYHLPDAESDEDEDFKEQTRLLKASIPFSVVGSNQLIEAKGKKVRGRLYPWGVVEVENPEHNDFLKLRTMLITHMQDLQEVTQDLHYENFRSERLKRGGRKVENEDMNKDQILLEKEAELRRMQEMIARMQAQMQMQMQGGDGDGGALGHHV

• Full Name: septin 2
• Calculated Molecular Weight: 41 kDa
• Observed Molecular Weight: 42-45 kDa
• GenBank Accession Number: BC014455
• Gene Symbol: Septin 2
• Gene ID (NCBI): 4735
• RRID: AB_2187016
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q15019
• 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

Septin 2, also named as DIFF6, KIAA0158, NEDD5, belongs to the septin family. It is a filament-forming cytoskeletal GTPase. Septin 2 is required for normal organization of the actin cytoskeleton. It plays a role in ciliogenesis and collective cell movements. Septin 2 is coordinated expression with Septin 6 and Septin 7. (PMID:21737677)

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	IF	Nat Genet; A transition zone complex regulates mammalian ciliogenesis and ciliary membrane composition.; Authors - Garcia-Gonzalo Francesc R FR
human	WB	Cell Res; Regulators of tubulin polyglutamylation control nuclear shape and cilium disassembly by balancing microtubule and actin assembly.; Authors - Lei Wang
human	WB	J Cell Biol; Septins suppress the release of vaccinia virus from infected cells.; Authors - Julia Pfanzelter
mouse	WB,IF	Cell Rep; Cdc42EP3/BORG2 and Septin Network Enables Mechano-transduction and the Emergence of Cancer-Associated Fibroblasts.; Authors - Fernando Calvo
human	CoIP	iScience; Contribution of septins to human platelet structure and function.; Authors - Oleg V Kim
canine	WB	J Cell Sci; Borg5/Cdc42EP1 restricts contractility and motility in epithelial MDCK cells; Authors - David Cohen

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Amaury (Verified Customer) (03-29-2022)

I performed immunofluorescence on K562. It gave me very nice immunolabelling. I used ice-cold methanol to permeabilize the cells.

• Applications: Immunofluorescence
• Cell Tissue Type: K562

Septin 2 Antibody Immunofluorescence validation ( dilution) in K562 (Cat no:60075-1-Ig)

FH

Stephen (Verified Customer) (01-31-2019)

works fine for WB in HeLa cells 10ug of protein lysate loaded onto 10% NuPAGE gel run I in 1XMOPS buffer blot transferred onto nitrocelluloseAntibody used at 1:2000 incubated overnight at 4 degrees t in 5% milk PBST solution.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: HeLa

Septin 2 Antibody Western Blot, validation (1:2000 dilution) in HeLa (Cat no:60075-1-Ig)