VCAM1 Recombinant Rabbit Monoclonal Antibody [SA05-04]
cat.: ET1601-18
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC(Intra)
Clonality: Monoclonal
Clone number: SA05-04
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 81 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human VCAM1 aa 690-739 / 739.
Positive control: C2C12 cell lysate, 3T3-L1 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, NIH/3T3 cell lysate, human spleen tissue, C2C12 cell lysate, C2C12.
Subcellular location: Membrane
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
  FC(Intra)
1:5,000-1:10,000
1:1,000
1:100
1:1,000-1:2,000
1-2μg/sample
1:1,000
Uniprot #: SwissProt: P19320 Human | P29533 Mouse | P29534 Rat
Alternative names: CD106 CD106 Antigen INCAM 100 INCAM-100 L1CAM MGC99561 V-CAM 1 Vascular Cell Adhesion Molecule 1 Vascular cell adhesion protein 1 VCAM 1 VCAM-1 VCAM1 VCAM1_HUMAN
Images
ET1601-18_1.jpg Fig1: Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: C2C12 cell lysate (20 µg/Lane)
Lane 2: 3T3-L1 cell lysate (20 µg/Lane)
Lane 3: Mouse spleen tissue lysate (20 µg/Lane)
Lane 4: Rat spleen tissue lysate (20 µg/Lane)

Predicted band size: 81 kDa
Observed band size: 100 kDa

Exposure time: 3 minutes; 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 (ET1601-18) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.
ET1601-18_2.jpg Fig2: Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/5,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: C2C12 cell lysate (20 µg/Lane)
Lane 3: RAW264.7 cell lysate (negative) (20 µg/Lane)
Lane 4: Mouse spleen tissue lysate (20 µg/Lane)
Lane 5: Rat spleen tissue lysate (20 µg/Lane)

Predicted band size: 81 kDa
Observed band size: 100 kDa

Exposure time: 24 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 (ET1601-18) at 1/5,000 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.
ET1601-18_3.jpg Fig3: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1601-18_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1601-18_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1601-18_6.jpg Fig6: VCAM1 was immunoprecipitated in 0.2mg C2C12 cell lysate with ET1601-18 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-18 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: C2C12 cell lysate (input)
Lane 2: Rabbit IgG instead of ET1601-18 in C2C12 cell lysate
Lane 3: ET1601-18 IP in C2C12 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 5 seconds; ECL: K1802
ET1601-18_7.jpg Fig7: Immunocytochemistry analysis of C2C12 cells labeling VCAM1 with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/1,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-VCAM1 antibody (ET1601-18) at 1/1,000 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 (M1305-2, 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.
ET1601-18_8.jpg Fig8: Flow cytometric analysis of C2C12 cells labeling VCAM1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-18, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.