MAPK11 antibody (N-Term)

Catalog No. ABIN2798348

Product Details anti-MAPK11 Antibody

Target: MAPK11 (Mitogen-Activated Protein Kinase 11 (MAPK11))

Binding Specificity: AA 1-30, N-Term

Reactivity: Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This MAPK11 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC)

Purpose: Rabbit Anti-Mouse Mapk11 (N-term) Antibody

Immunogen: This Mouse Mapk11 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 1-30 amino acids from the N-terminal region of mouse Mapk11.

Isotype: Ig Fraction

Application Details

Application Notes: Western Blot, Immunohistochemistry
Recommended Dilutions
WB: 1:1000, IHC: 1:10-50Western blot analysis of lysate from mouse NIH/3T3 cell line, using Mouse Mapk11 Antibody (N-term) for immunohistochemistry. Clinical relevance has not been evaluated.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #100108 (Western Blotting)

by: Instituto de Parasitología y Biomedicina López-Neyra

No.: #100108

Date: 12/23/2016

Antigen: MAPK11

Lot Number: RAY8110713

Method validated: Western Blotting

Positive Control: primary mouse microglia

Negative Control: siRNA knock-down of MAPK11 in primary mouse microglia

Notes: Passed. ABIN2798348 specifically recognizes Mapk11 in primary mouse microglia lysates.

Validation Images

Primary mouse microglia were transfected with Mapk11 siRNA and Mapk11 was revealed using ABIN2798348 as described in the protocol section (lanes 1, 2, and 3). Lysates from cells transfected with scrambled siRNA (lane 4) and untransfected cells (lane 5) served as negative controls.

Full Methods

Primary Antibody: ABIN2798348

Secondary Antibody: anti-rabbit HRP-conjugated antibodies (DakoCytomation, P0448, lot 00055814)

Full Protocol:

• Primary mouse microglia are grown in DMEM (Invitrogen), supplemented with 10% foetal bovine serum (Gibco), 10% horse serum and penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 12-well dish.
• Transfect cells with 0.05nmol mouse MAPK11 Silencer Select siRNA (ThermoFisher Scientific, Sample ID ASO10943, ASO1098J and ASO109CX, lot AMO04BXD) using Lipofectamine 3000 (Invitrogen) following the manufacturer´s instructions.
• Lyse cells in 25µl per well cold lysis buffer (10mM Tris-HCl pH 8.0, 150mM NaCl, 1% Nonidet-P40, 1mM EDTA, 10mM NaF, 1mM Na₃VO₄, protease inhibitors (Sigma).
• Determine total protein content of the lysates using Bradford assay (Bio-Rad, 500-0006, lot 111832).
• Denature 5µg total protein for 5min at 95°C in 5µl Laemmli SDS sample buffer and subsequently separate them on a denaturing freshly cast 10% SDS-PAGE for 1h at 140V.
• Transfer proteins onto PVDF membrane (Pall Life Sciences, 75696G, lot TO3225) with a semi-dry Western blotting system for 50min at 60mA.
• Block the membrane with TBST (TBS, 0.1% Tween) containing 5% milk ON at 4°C.
• Incubation with primary rabbit anti-MAPK11 antibody (antibodies-online, ABIN2798348, lot RAY8110713) diluted 1:500 in TBST containing 5% milk for 1h at RT.
• Wash membrane 5x 5min with TBST.
• Incubation with anti-rabbit horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0448, lot 00055814) diluted 1:2000 in TBST containing 5% milk for 1h at RT.
• Wash membrane 5x 5min with TBST and 3x 5min with TBS.
• Reveal protein bands using ECL Prime Western Blotting Detection Reagent (GE Healthcare, RPN2232, lot 9547481) with Curix RP2 Plus medical X-ray films (AGFA Health Care, ENKMV, lot 79040074) on an AGFA Health Care Curix60-developer.
• Strip membranes RestoreTM Western Blot Stripping Buffer (Thermo Scientific, 21059, lot NA 164269) and subsequently incubate with a GAPDH loading control antibody (Sigma, G9545, lot 049K4787) diluted 1:10000 and with anti-rabbit horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0448, lot 00055814) diluted 1:2000.
• Wash and reveal the protein bands using self-made ECL Western Blotting Detection Reagent with Curix RP2 Plus medical X-ray films (AGFA Health Care, ENKMV, lot 79040074) on an AGFA Health Care Curix60-developer.

Experimental Notes: The MAPK11 antibody ABIN2798348 reveals a protein of the expected molecular weight of murine Mapk11 in lysates of primary mouse microglia. The protein band’s intensity decreases upon siRNA knockdown.

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Storage: 4 °C,-20 °C

Storage Comment: 2-8°C (short-term), -20°C (long-term)

Target Details for MAPK11

Target: MAPK11 (Mitogen-Activated Protein Kinase 11 (MAPK11))

Alternative Name: Mapk11 (MAPK11 Products)

Synonyms: ATMPK11 antibody, F22L4.10 antibody, F22L4_10 antibody, MAP kinase 11 antibody, zgc:86905 antibody, P38B antibody, P38BETA2 antibody, PRKM11 antibody, SAPK2 antibody, SAPK2B antibody, p38-2 antibody, p38Beta antibody, P38b antibody, Prkm11 antibody, Sapk2 antibody, Sapk2b antibody, p38beta antibody, p38beta2 antibody, p38b antibody, sapk2 antibody, prkm11 antibody, sapk2b antibody, mitogen-activated protein kinase 11 antibody, MAP kinase 11 antibody, mapk11 antibody, LOC9328118 antibody, MPK11 antibody, MAPK11 antibody, Mapk11 antibody, PTRG_02565 antibody

Background:

Target Description: Kinase involved in a signal transduction pathway that is activated by changes in the osmolarity of the extracellular environment, by cytokines, or by environmental stress. Phosphorylates preferentially transcription factor ATF2 (By similarity).

Gene Symbol: Mapk11

Molecular Weight: 41397 Da

Gene ID: 19094

UniProt: Q9WUI1

Pathways: MAPK Signaling, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Response to Water Deprivation, Regulation of Muscle Cell Differentiation, ER-Nucleus Signaling, Hepatitis C, Toll-Like Receptors Cascades, Signaling Events mediated by VEGFR1 and VEGFR2, Thromboxane A2 Receptor Signaling, BCR Signaling, S100 Proteins