R1308-3

p38 alpha / MAPK14 Rabbit Polyclonal Antibody

cat.: R1308-3

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 41 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within C-terminal human MAPK14.
Positive control:	HeLa cell lysate, Jurkat cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, C6 cell lysate, HeLa, human kidney tissue, human stomach tissue, mouse kidney tissue, mouse stomach tissue, rat kidney tissue, rat stomach tissue.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000 1:100-1:200 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: Q16539 Human \| P47811 Mouse \| P70618 Rat
Alternative names:	CSAID Binding Protein 1 CSAID binding protein CSAID-binding protein Csaids binding protein CSBP 1 CSBP 2 CSBP CSBP1 CSBP2 CSPB1 Cytokine suppressive anti-inflammatory drug-binding protein EXIP MAP kinase 14 MAP kinase MXI2 MAP kinase p38 alpha MAPK 14 MAPK14 MAX interacting protein 2 MAX-interacting protein 2 Mitogen Activated Protein Kinase 14 Mitogen activated protein kinase p38 alpha Mitogen-activated protein kinase 14 Mitogen-activated protein kinase p38 alpha MK14_HUMAN Mxi 2 MXI2 p38 ALPHA p38 p38 MAP kinase p38 MAPK p38 mitogen activated protein kinase p38ALPHA p38alpha Exip PRKM14 PRKM15 RK SAPK2A

Images

Fig1: Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: MCF7 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: PC-12 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 45 kDa

Exposure time: 25 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 (R1308-3) at 1/5,000 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.

Fig2: Immunocytochemistry analysis of HeLa cells untreated / treated with UV for 30 minutes then recover for 30 minutes labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/100 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-p38 alpha / MAPK14 antibody (R1308-3) at 1/100 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-p38 alpha / MAPK14 antibody (R1308-3) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1308-3) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig9: Flow cytometric analysis of HeLa cells labeling p38 alpha / MAPK14.

Cells were fixed and permeabilized. Then stained with the primary antibody (R1308-3, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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