HSPA1L Mouse Monoclonal Antibody [7D3]
cat.: EM1801-16
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: 7D3
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 70 kDa
Isotype: IgG1
Immunogen: Recombinant protein with Human HSPA1L aa 335-639 / 641.
Positive control: Mouse testis tissue lysate, rat testis tissue lysate, A549 cell lysates, HeLa, human testis tissue, mouse testis tissue, rat testis tissue.
Subcellular location: Cytosol, Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC

1:2,000
1:100
1:1,000
1:1,000
Uniprot #: SwissProt: P34931 Human | P16627 Mouse | P55063 Rat
Alternative names: Heat shock 70 kDa protein 1 Hom Heat shock 70 kDa protein 1 like Heat shock 70 kDa protein 1-Hom Heat shock 70 kDa protein 1-like Heat shock 70 kDa protein 1L Heat shock 70kD protein like 1 HS71L_HUMAN HSP70 1L HSP70 HOM HSP70-Hom HSPA1L hum70t Spermatid specific heat shock protein 70
Images
EM1801-16_1.jpg Fig1: Western blot analysis of HSPA1L on different lysates with Mouse anti-HSPA1L antibody (EM1801-16) at 1/2,000 dilution.

Lane 1: Mouse testis tissue lysate
Lane 2: Rat testis tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 70 kDa
Observed band size: 70 kDa

Exposure time: 24 seconds;

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 (EM1801-16) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.
EM1801-16_2.jpg Fig2: Western blot analysis of HSPA1L on A549 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1/500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Lane 1: Anti-HSPA1L Antibody.
Lane 2: Anti-HSPA1L Antibody, preincubated with immunization protein.
EM1801-16_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling HSPA1L with Mouse anti-HSPA1L antibody (EM1801-16) at 1/100 dilution.

Cells were fixed in 100% precooled methanol 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 Mouse anti-HSPA1L antibody (EM1801-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
EM1801-16_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-HSPA1L antibody (EM1801-16) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-16) 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.
EM1801-16_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-HSPA1L antibody (EM1801-16) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-16) 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.
EM1801-16_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-HSPA1L antibody (EM1801-16) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-16) 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.
EM1801-16_7.jpg Fig7: Flow cytometric analysis of HeLa cells labeling HSPA1L.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1801-16, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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