ET1704-49

Annexin A2 Recombinant Rabbit Monoclonal Antibody [JA42-30]

cat.: ET1704-49

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, IF-Tissue, FC
Clonality:	Monoclonal
Clone number:	JA42-30

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 39 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Annexin A2 aa 26-75 / 339.
Positive control:	A549 cell lysate, HeLa cell lysate, K-562 cell lysate, 293T cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Hela, human liver tissue, human colon carcinoma tissue, human prostate tissue, K562.
Subcellular location:	Basement membrane. Melanosome.
Recommended Dilutions: WB IHC-P IF-Cell IF-Tissue FC	1:1,000 1:50-1:200 1:100-1:200 1:100-1:200 1:50-1:200
Uniprot #:	SwissProt: P07355 Human \| P07356 Mouse \| Q07936 Rat
Alternative names:	Annexin A2 Annexin II Annexin II, heavy chain Annexin-2 ANX 2 ANX2 ANX2L4 ANXA2 ANXA2_HUMAN arylsulfatase B CAL1H Calpactin I heavy chain calpactin I heavy polypeptide (p36) Calpactin I heavy polypeptide Calpactin-1 heavy chain chromobindin 8 Chromobindin-8 Epididymis secretory protein Li 270 HEL S 270 LIP2 Lipocortin II LPC2 LPC2D p36 P36 protein PAP-IV Placental anticoagulant protein IV Protein I

Images

Fig1: Western blot analysis of Annexin A2 on different lysates with Rabbit anti-Annexin A2 antibody (ET1704-49) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD Annexin A2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 34 kDa

Exposure time: 12 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 (ET1704-49) at 1/1,000 dilution was used in 5% NFDM/TBST 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: Western blot analysis of Annexin A2 on different lysates with Rabbit anti-Annexin A2 antibody (ET1704-49) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: 293T cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 35 kDa

Exposure time: 4 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 (ET1704-49) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: ICC staining of Annexin A2 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-49, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Annexin A2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-49, 1/50) for 30 minutes 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 human colon carcinoma tissue using anti-Annexin A2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-49, 1/50) for 30 minutes 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 human prostate tissue using anti-Annexin A2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-49, 1/50) for 30 minutes 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: Flow cytometric analysis of Annexin A2 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-49, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.