Anti-HMGA1a / HMGA1b 抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody (AB4078)

IHC image of ab4078 staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab4078, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-HMGA1a / HMGA1b antibody (AB4078)

Immunofluoresence on MCF-7 cells transfected with human HMGA1a-GFP using ab4078.

A : Hoechst stain
B : Phasecontrast
C : TexasRed HMG-I/HMG-Y (ab4078)
D : HMGA1a-GFP
E : Merge

This image is courtesy of Robert Hock

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody (AB4078)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling HMGA1 with ab315350 at 1/2000 (0.251 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human testis in (A) ab315350 and (B) ab4078.
The image on (A) ab315350 is applied with Anti-HMGA1 antibody at 1/2000 dilution and (B) ab4078 is applied with Anti-HMGA1 antibody at 1/500 dilution.
The section was incubated with ab315350 or ab4078 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Collaborator

Western blot - Anti-HMGA1a / HMGA1b antibody (AB4078)

Western blot using ab4078.

Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a-, hHMGA1b-, hHMGA2- or NLS- (nuclear localisation signal) GFP fusion proteins.

ab4078 was diluted 1/1000 in 5% milk/TBS and incubated for 1 hr at room temperature. Secondary HRP anti-rabbit antibody was diluted 1/10000 in 5% milk/TBS and incubated for 1 hr at room temperature.

Lane 1 : hHMGA1a-GFP (ab4078)
Lane 2 : hHMGA1b-GFP (ab4078)
Lane 3 : hHMGA2-GFP (ab4078)
Lane 4 : NLS-GFP (ab4078)
Lane 5 : hHMGA1a-GFP (anti-GFP)
Lane 6 : hHMGA1b-GFP (anti-GFP)
Lane 7 : hHMGA2-GFP (anti-GFP)
Lane 8 : NLS-GFP (anti-GFP)

The detected size is that expected for the fusion of an 11kD and 26kD protein (GFP).

Lanes 1 - 4:

Western blot - Anti-HMGA1a / HMGA1b antibody (ab4078) at 1/1000 dilution

Lanes 5 - 8:

Anti-GFP

Lane 1:

Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a GFP fusion proteins

Lanes 2 and 6:

Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1b-GFP fusion proteins

Lanes 3 and 7:

Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA2-GFP fusion proteins

Lanes 4 and 8:

Whole cell lysate from HepG2 cells transfected with plasmids coding for NLS-GFP fusion proteins

Lane 5:

Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a-GFP fusion proteins

Secondary

All lanes:

Anti-rabbit HRP at 1/10000 dilution

Predicted band size: 12 kDa

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This image is courtesy of Monika Harrer

• WB

CiteAb

Western blot - Anti-HMGA1a / HMGA1b antibody (AB4078)

HMGA1a / HMGA1b western blot using anti-HMGA1a / HMGA1b antibody ab4078. Publication image and figure legend from Zhang, H., Yang, J., et al., 2018, Oncotarget, PubMed 29581847.

ab4078 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab4078 please see the product overview.

Consequences of HMGA1 silencing in airway BC on the expression of genes associated with airway remodeling in COPD(A) Effect of HMGA1 silencing on expression of squamous, epithelial-mesenchymal transition (EMT), inflammation and senescence-related genes in BC differentiating on ALI. Gene expression was assessed by TaqMan at ALI day 14. Shown are squamous-related genes keratin 6B (KRT6B), involucrin (IVL) and stratifin (SFN); EMT-related genes vimentin (VIM), collagen type I α1 (COL1A1), and matrix metallopeptidase 2 (MMP2); and inflammation-related genes interleukin 1α (IL1A), interleukin 1β (IL1B), interleukin 6 (IL6), interleukin 8 (IL8), prostaglandin-endoperoxide synthase 2 (cyclooxygenase 2 or PTGS2) and senescence-related gene cyclin dependent kinase inhibitor 1A (CDKN1A). Data was normalized to 18S rRNA. Gene expression in control siRNA transfected samples was normalized to 1. n = 6, each group. For A and B, LAE BC from Lonza, see Methods, BC-1 and ALI-1. (B) Immunofluorescence assessment of protein expression. Purified normal SAE BC were transfected by control and HMGA1 siRNAs and the capacity of these cells to differentiate were assessed in ALI culture. At ALI day 14, the membranes were paraffin embedded and sectioned. The cells grown on the membranes were then characterized by immunofluorescences using squamous cell marker IVL (green) and EMT marker VIM (red). Dashed line indicates the ALI membrane. See Methods, BC-2 and ALI-2. (C) Western analysis. HMGA1 was silenced by HMGA1 siRNA transfection in BC. At ALI day 14, protein was harvested and Western analysis was performed using SCGB1A1, DNAI1, keratin 6 (KRT6), IVL, VIM, cadherin 1 (CDH1, also known as E-cadherin), cadherin 2 (CDH2, also known as N-cadherin), and HMGA1 antibodies, respectively. GAPDH was used as a loading control. LAE BC from Lonza, see Methods, BC-1 and ALI-1.

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