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TNF alpha Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# R1203-1

TNF alpha Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • ELISA

Reactivity

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

TNF alpha Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within mouse TNF alpha aa 40-80 and aa 170-210.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell, ELISA

Molecular Weight

Predicted band size: 26 kDa

Positive Control

RAW264.7 treated with 100ng/mL LPS for 7 hours add 300ng/mL BFA for last 3 hours cell lysate, Recombinant murine TNF alpha.

Conjugation

unconjugated

RRID

AB_3073194

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:10,000

  • IF-Cell

  • 1:500

  • ELISA

  • 1:10,000-20,000

Target

Function

TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein. Membrane bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer. Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells and macrophages. TNF-α plays a key regulatory role in infammation and host defense against bacterial infection, notably Mycobacterium tuberculosis. The antibody detects active recombinant murine TNF alpha. The detective sensitivity by Western Blot is 5ng.

Background References

1. Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.

2. Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.

3. Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.

4. Brennan, F.M. and McInnes, I.B. (2008) J Clin Invest 118, 3537-45.

Sequence Similarity

Belongs to the tumor necrosis factor family.

Post-translational Modification

The membrane-bound form is further proteolytically processed by SPPL2A or SPPL2B through regulated intramembrane proteolysis producing TNF intracellular domains (ICD1 and ICD2) released in the cytosol and TNF C-domain 1 and C-domain 2 secreted into the extracellular space (By similarity). The soluble form derives from the membrane form by proteolytic processing.; The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1 (By similarity).; O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.

Subcellular Location

Secreted, membrane, cytoplasm.

UNIPROT

SwissProt: P06804 Mouse

Synonyms

APC1 antibody

APC1 protein antibody

Cachectin antibody

DIF antibody

Differentiation inducing factor antibody

Macrophage cytotoxic factor antibody

Tnf antibody

TNF superfamily, member 2 antibody

TNF, macrophage derived antibody

TNF, monocyte derived antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of TNF alpha on different lysates with Rabbit anti-TNF alpha antibody (<a href="/products/R1203-1" style="font-weight: bold;text-decoration: underline;">R1203-1</a>) at 1/10,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for last 3 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 17/26/33 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/R1203-1" style="font-weight: bold;text-decoration: underline;">R1203-1</a>) at 1/10,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of TNF alpha on different lysates with Rabbit anti-TNF alpha antibody (R1203-1) at 1/10,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for last 3 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 17/26/33 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (R1203-1) at 1/10,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for last 3 hours labeling TNF alpha with Rabbit anti-TNF alpha antibody (<a href="/products/R1203-1" style="font-weight: bold;text-decoration: underline;">R1203-1</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TNF alpha antibody (<a href="/products/R1203-1" style="font-weight: bold;text-decoration: underline;">R1203-1</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for last 3 hours labeling TNF alpha with Rabbit anti-TNF alpha antibody (R1203-1) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TNF alpha antibody (R1203-1) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Western Blot analysis of recombinant murine TNF-alpha using rabbit TNF alpha antibody.<br />Western blot analysis of TNF alpha on recombinant protein lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/R1203-1" style="font-weight: bold;text-decoration: underline;">R1203-1</a>, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.

    Western Blot analysis of recombinant murine TNF-alpha using rabbit TNF alpha antibody.
    Western blot analysis of TNF alpha on recombinant protein lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (R1203-1, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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TNF alpha Recombinant Rabbit Monoclonal Antibody [PS01-07]

Application: WB

Reactivity: Human,Mouse

Conjugate: unconjugated